Research Solutions - GRiSPgrisp.pt/docs/grisp2016.pdfanalysis, restriction enzyme digest analysis, Northern Blotting, etc. NOTE: GRiSP Research Solutions recommends using GRS Agarose

Research Solutions

GRiSP Research Solutions would like to welcome you to our 2016 catalogue

GRiSP is a Portuguese company, founded in 2008, born from the will of its founders to support the research market with high quality reagents and kits, in the areas of molecular biology, biochemistry, genetics, and related.

Dedicated to developing, producing and commercializing, our team is highly motivated to bring innovative, as well as known products to the market with high quality, and competitive pricing, making it easier for researchers to achieve great results, with minimum cost and effort.

We believe this catalogue will help you access a comprehensive range of products, focusing on DNA Electrophoresis, Nucleic Acid Purification, PCR, qPCR, RNA Research, Cloning, Protein Research, Cell Biology and Culture Media.

Feel free to contact us with your questions, suggestions, or any other situation.

GRiSP Team

Terms and Conditions:

GRiSP, Lda. reserves the right to change prices as well as terms and conditions, without previous notice.GRiSP, Lda. is committed to the quality of its products. However, if a product is proven to be defective (e.g. caused by transport), returns are accepted within 10 days after receiving the product. Products ordered by mistake can be returned within 10 days after receiving the product, if unused and not damaged, and transport costs for adequate ship-ping conditions are to be assumed by the customer.Payment terms are 30 days, and VAT is not included in the prices shown (only applicable for Portugal).

Placing an order:

GRiSP accepts orders via E-mail or Fax.

VAT NR: PT508573920

Payment Options

GRiSP accepts payment via cheque or bank transfer. For payment by cheque, please use the postal adress mentioned above. For Bank Transfer use the following data:

Delivery time

Portugal: Courier; 1-2 working days after order (except in case of stock rupture)Europe: Courier; 2-5 working days after order (except in case of stock rupture)For Express or Other Countries: please contact us for an estimation of the shipping time

Postal Address:

GRiSP, Lda.Rua Alfredo Allen, 4554200-135 PortoPortugal

Please use one of the following contacts:

Email: [email protected]: +351 220 301 599Fax: +351 220 301 597

Grisp, LdaIBAN: PT50 0046 0109 0060 0193 9380 2BIC/SWIFT: CRBNPTPLBank: Banco Popular Portugal, SA

Account:

Research Solutions

Product CategoriesDNA Electrophoresis

Nucleic Acid Purification

DNA Amplification

RNA Research

Cloning

Culture Media

Protein Research

Cell Biology

5

13

37

47

53

61

65

73

DNA Electrophoresis

Buffers

SGTB Agarose Electrophoresis Buffer

TAE Buffer

TBE Buffer

GRS DNA Blue Loading Buffer (6x)

Agaroses

GRS Agarose LE

GRS Agarose LE (in tablets)

GRS Agarose LMT

GRS Agarose S-LMT

06

07

07

07

08

08

09

09

01DNA Ladders

GRS Ladder 50bp

GRS Ladder 100bp

GRS Ladder 1Kb

GRS LowRange Ladder

GRS HighRange Ladder

GRS Universal Ladder

10

10

10

11

11

11

6

Fig 1/ Comparison of SGTB vs TBE: A 50bp DNA ladder (200bp, 500bp, and 1kb are more intense for identification) was resolved by 2% agarose electro-phoresis, using a standard agarose (LE), on a 10cm length minigel until the 200bp band migrated 45mm. Initial temperatures were 20ºC and final temperatures were 38ºC and 29ºC, resp.

Fig 2/ DNA marker (100bp, 250bp, 500bp, 750bp, 1000bp, 1500bp, and 2000bp) re-solved in 25 min at 200V using 1,5% standard agarose gel electrophoresis

Fig 3/ Comparison of SGTB vs TBE: relationship between Relative distances and DNA size. A 50-bp DNA ladder was separated using 2% standard agarose. Since tracking dyes migrate differently in gels prepared with SGTB, Relative mobility was determined relatively to the 200bp band. As can be clearly seen, using SGTB results in a much better separation of DNA fragments in the whole range of 500bp to 50bp than with using TBE, as distances between similar sizes are greater.

500bp

SGTB TBE

500bp

200bp

SGTB Agarose Electrophoresis BufferSGTB is a NEW buffer ideal for Agarose Elec-trophoresis of DNA fragments of 100bp to 1000bpTime Saving

SGTB allows much FASTER RUNS.

Unlike TAE and TBE gels, which tolerate only about 110V (mini-gel), the low-conductance electrophoretic buffer SGTB allows for the application of high voltage (200-300V) without generating a lot of heat, so your gel won’t melt.

High Resolution

DNA agarose gel electrophoresis using SGTB results in very sharp bands. Faster Runs result in less fuzzy bands.

Better Separation

Using SGTB results in larger differences in relative mobility (Rf), espe-cially between smaller DNA fragments.

Stronger and Clearer

Gels made with SGTB are also CLEARER, increasing detection sensitiv-ity; and STRONGER, making them excelent for blotting.

If you already get the separation you need with TBE/TAE, you can get the same resolution with SGTB using less agarose. With SGTB you save TIME and MONEY

Reference Description Quantity

GB01.0120 SGTB Electrophoresis Buffer 1L 20X



Ideal for separation of PCR products

Very fast

Saves ~25% Agarose

DNA Electrophoresis / Buffers

www.grisp.ptalways here for you

www.grisp.pt

7

TAE Buffer (10x)(Tris-Acetate-EDTA, 10x concentrated buffer)Tris-acetate-EDTA (TAE) buffer is the most common buffer used for agarose gel electrophoresis in the analyses of DNA samples. The choice of the electrophoresis buffer to be used depends on the nature of the sample and downstream applications. TAE buffer is recommend-ed for high resolution of large DNA fragments (>3kb) or supercoiled DNA, however, it can also be used for non-denaturing RNA agarose electrophoresis.

Features

TAE Buffer (10x) is a 0.2µm filtered aqueous solution of 400mM Tris, 200mM glacial acetic acid, and 10mM EDTA prepared with ultrapure water. It is supplied in a 1L plastic bottle or in a 5L handy Bag-in-Box equipped with a two-position spigot that offers push button dispensing.

TBE Buffer (10x)(Tris-Borate-EDTA, 10x concentrated buffer)Tris-Borate-EDTA (TBE) buffer can be used for both agarose and polyacryl-amide gel electrophoresis. The choice of the electrophoresis buffer to be used depends on the nature of the sample and downstream applications. TBE buffer is recommended for high resolution of small DNA fragments (<1,5kb), e.g. PCR products or products of restriction enzyme digestion, however, it is also suitable for RNA polyacrylamide electrophoresis.

Features

TBE Buffer (10x) is a 0.2µm filtered aqueous solution of 0,89 M Tris, 0,89 M Boric Acid, and 0,02M EDTA prepared with ultrapure water. It is supplied in a 1L plastic bottle or in a 5L handy Bag-in-Box equipped with a two-position spigot that offers push button dispensing.

GRS DNA Loading Buffer Blue (6X)DNA Loading Buffer Blue (6x) is a pre-mixed Loading Buffer for Agarose and Polyacrylamide gel electrophoresis, containing Bromophenol Blue and Xylene Cyanol FF as tracking dyes. EDTA is included to chelate Magnesium (up to 10 mM), stopping enzymatic reac-tions.

Dye Migration (1% agarose gels):

Xylene Cyanol FF

TAE: ~ 6kb

TBE: ~ 4kb

SGTB: ~ 10kb

Bromophenol Blue

TAE: ~ 500bp

TBE: ~ 400bp

SGTB: ~ 125bp

Composition:

10 mM Tris-HCl pH 7,6

60 mM EDTA

0,03% Bromophenol Blue

0,03% Xylene Cyanol FF

60% Glycerol


GB11.0110 TAE Buffer 10x 1L

GB11.0510 TAE Buffer 10x 5L


GB12.0110 TBE Buffer 10x 1L

GB12.0510 TBE Buffer 10x 5L


GLB01.0001 DNA Loading Buffer Blue (6x)

1ml

GLB01.0501 DNA Loading Buffer Blue (6x)

5x 1ml

DNA Electrophoresis / Buffers

8

GRS Agarose LEStandard Agarose for routine analytical and preparative DNA gelGRS Agarose LE is suitable for all analytical and prepara-tive electrophoresis of nucleic acids in routine gel electro-phoresis. Depending on the concentration of GRS Agarose LE, the size range of nucleic acid separation wil l vary between 100bp and 15 kb. Because of its low EEO, DNA will have a high electrophoretic mobility.

GRS Agarose LE is useful for a broad range of applications: PCR product analysis, restriction enzyme digest analysis, Northern Blotting, etc.

NOTE: GRiSP Research Solutions recommends using GRS Agarose LE with SGTB DNA Electrophoresis Buffer. Using SGTB Electrophoresis Buffer you can run gels faster, with sharper bands and even save up to 25% Agarose with the same or better resolution!

GRS Agarose LE in tabletsStandard Agarose in tablets, for routine analytical and preparative DNA gel electro-phoresisGRS Agarose LE in tablets is suitable for all analytical and prepar-ative electrophoresis of nucleic acids in routine gel electrophore-sis. Depending on the concentration of GRS Agarose LE, the size range of nucleic acid separation will vary between 100bp and 15 kb. Because of its low EEO, DNA will have a high electrophoretic mobility. GRS Agarose LE in tablets is useful for a broad range of applications: PCR product analysis, restriction enzyme digest analysis, Northern Blot-ting, etc.

NOTE: GRiSP Research Solutions recommends using GRS Agarose LE in tablets with SGTB DNA Electrophoresis Buffer. Using SGTB Electro-phoresis Buffer you can run gels faster, with sharper bands and even save up to 25% Agarose with the same or better resolution!

Range: 100bp – 15kb



GA110.0500 GRS Agarose LE 500g

DNase/RNase/Protease Free


GA110.100T GRS Agarose LE in tablets 100x 0.5g

GA110.200T GRS Agarose LE in tablets 200x 0.5g

DNA Electrophoresis / Agaroses


www.grisp.pt

9

GRS Agarose LMTLow melting Temperature Agarose, for DNA/RNA/Proteins & in gel reactionsGRS Agarose LMT is a high purity low melting agarose, excellent for separation of RNA and DNA fragments as well as isolation of genomic DNA and proteins. Optimum separation of DNA in the 100bp to 10kb range will be obtained using GRS Agarose LMT at concentra-tions between 0,75% and 1,75%. The low gelling temperature (24°C - 28°C) facilitates quantitative recovery of nucleic acids, the inclusion of thermo-labile substances such as living cells, and allows enz ymatic manipulation (enzymatic digest, ligation, nick translation, etc.) of DNA in melted agarose without cutting slices. For optimal resolution and results, it is highly recommended to keep the gels at +4°C for at least 1 hour before running the gel.

GRS Agarose S-LMTLow melting, fine resolution of small DNA fragmentsGRiSP Agarose S-LMT has been designed for electrophoretic analysis of small nucleic acids. Fine resolution of small DNA fragments or PCR products in the 50bp to 1000bp range can be obtained. It can also be used for genotyping, allele sizing and short tandem repeat analysis. GRS Agarose S-LMT works as a molecular screen and has twice the resolution of the finest sieving agaroses. It can discriminate between fragments which differ by only a fewbp in length and thus, it is able to compete with acrylamide gels, being much easier to handle as the latter. GRS Agarose S-LMT 3% to 6% gels give similar results to Poly-acrylamide 6% to 8% gels. For efficient resolution, the concentration of GRS Agarose S-LMT should be adjusted according to the range of band sizes analyzed, and the gel running buffer used. The low gelling temperature (<35ºC) facilitates quantitative recovery of nucleic acids, the inclusion of thermoslabile substances such as living cells, and allows enzymatic manipulation (enzymatic digest, ligation, nick translation) of DNA in melted agarose.


GA115.0050 GRS Agarose LMT 50g


GA116.0100 GRS Agarose S-LMT 100g

Range: 50bp – 1kb


DNA Electrophoresis / Agaroses

10

8000

5000

3000

1500

500

GRS Ladder 50bpRecommended Loading 2-5 μL/lane (50 μg allow for ~ 80-200 lanes)GRS DNA Ladder 50bp is a ready-to-use DNA Marker suitable for sizing linear double-stranded DNA fragments from 50bp-1000bp, and composed of 13 linear chromatography-purified individual DNA frag-ments. All bands (except 250bp and 500bp) are supplied at approxi-mately 40ng/5μL. The 250bp and 500bp bands have increased inten-sity relative to the other bands on an ethidium bromide-stained agarose gel.

GRS Ladder 100bpRecommended Loading 2-5 μL/lane (50 μg allow for ~ 100-250 lanes)GRS Ladder 100bp is a ready-to-use DNA Marker suitable for sizing linear double-stranded DNA fragments from 100bp-1500bp, and composed of 11 chromatography-purified individual DNA fragments. All bands (except 500bp) are supplied at approximately 40ng/5μL. The 500bp band has increased intensity relative to the other bands on an ethidium bromide-stained agarose gel.

bp

3% Agarose, 0.5x TAE, 50V - 1h minigel - 5μL/lane

1000800600

400

250

150

50

900700

500

300

200

100Reference Description Quantity

GL031.0050 GRS Ladder 50bp 50ug

GL031.5050 GRS Ladder 50bp 5x 50ug


GL041.0050 GRS Ladder 100bp 50ug

GL041.5050 GRS Ladder 100bp 5x 50ug

bp

1.7% Agarose, 0.5x TBE, 50V - 1h minigel - 5μL/lane

1500

900

700

500

300

100

1000800

600

400

200

GRS Ladder 1kbRecommended Loading 2-5 μL/lane (50 μg allow for ~ 100-250 lanes)GRS Ladder 1kb is a ready-to-use DNA Marker suitable for sizing linear double-stranded DNA fragments from 500bp-10kb, and composed of 10 chromatography-purified individual DNA fragments. All bands (except 2kb and 5kb) are supplied at approximately 40ng/5μL. The 2kb and 5kb bands have increased intensity relative to the other bands on an ethidium bromide-stained agarose gel.

1% Agarose, 1x TAE, 70V - 45min minigel - 5μL/lane

bp

10000

6000

4000

2000

1000


GL051.0050 GRS Ladder 1kb 50ug

GL051.5050 GRS Ladder 1kb 5x 50ug

DNA Electrophoresis / DNA Ladders


www.grisp.pt

11

1.7% Agarose, 0.5x TBE, 50V - 1h minigel - 5μL/lane

1% Agarose, 1x TAE, 70V - 45min minigel - 5μL/lane

GRS Low Range LadderRecommended Loading 2-5 μL/lane (50 μg allow for ~ 100-250 lanes)GRS Low Range Ladder is a ready-to-use DNA Marker suitable for sizing linear double-stranded DNA fragments from 25bp-700bp, and composed of 10 linear chromatography-purified individual DNA frag-ments. All bands (except 100bp and 300bp) are supplied at approxi-mately 40ng/5μL. The 100bp and 300bp bands have increased intensity relative to the other bands on an ethidium bromide-stained agarose gel.

bp

3% Agarose, 0.5x TAE, 50V - 1h minigel - 5μL/lane

700

400

200

100

50

500

300

150

75

25


GL011.0050 GRS Low Range Ladder 50ug

GL011.5050 GRS Low Range Ladder 5x 50ug


GL021.0050 GRS High Range Ladder 50ug

GL021.5050 GRS High Range Ladder 5x 50ug


GL061.0050 GRS Universal Ladder 50ug

GL061.5050 GRS Universal Ladder 5x 50ug

GRS High Range LadderRecommended Loading 2-5 μL/lane (50 μg allow for ~ 120-300 lanes)GRS High Range Ladder is a ready-to-use DNA Marker suitable for sizing linear double-stranded DNA fragments from 250bp-15Kb, and composed of 7 linear chromatography-purified individual DNA fragments. All bands (except 2500bp) are supplied at approximately 50ng/5μL. The 2500bp band has increased intensity relative to the other bands on an ethidium bromide-stained agarose gel.

GRS Universal LadderRecommended Loading 2-5 μL/lane (50 μg allow for ~ 70-180 lanes)GRS Universal Ladder is a ready-to-use DNA Marker suitable for sizing linear double-stranded DNA fragments from 100bp-10kb, and composed of 15 chromatography-purified individual DNA fragments. All bands (except 500bp and 3kb) are supplied at approximately 40ng/5μL. The 500bp and 3kb bands have increased intensity relative to the other bands on an ethidium bromide-stained agarose gel.

bp

10000

6000

4000

2000

1000

500

300

100

8000

5000

3000

1500

700

400

200

1% Agarose, 1x TAE, 70V - 1h minigel - 5μL/lane

1% Agarose, 1x TAE, 70V - 45 min minigel - 5μL/lane

15000

7500

2500

250

10000

5000

1000

bp

DNA Ladders

Nucleic Acid Purification

PCR Purification & Gel Extraction

GRS PCR & Gel Band Purification Kit

Exo/SAP Go – PCR Purification Kit

Cut&Spin Gel Extraction Columns

Xtractor Gel Band Cutter

Dye Terminator Clean-Up

GRS Dye Terminator Removal Resin

gDNA Purification

GRS Genomic DNA Kit – Blood & Cultured Cells

GRS Genomic DNA Kit – Tissue

GRS Genomic DNA Kit – Plant

GRS Genomic DNA Kit – Bacteria

GRS Genomic DNA Kit – BroadRange

GRS Pure DNA Kit

GRS Genomic DNA Kit - Card

RNA Purification

GRS Total RNA Kit – Blood & Cultured Cells

GRS Total RNA Kit – Tissue

GRS Total RNA Kit – Plant

GRS Total RNA Kit – Bacteria

GRS Total RNA Kit – Yeast & Fungus

16

17

18

18

19

20

20

21

21

22

22

23

24

24

25

25

26

02GRS Pure RNA Kit

tripleXtractor directRNA Kit

GRS microRNA Kit

DNA & RNA Purification

tripleXtractor reagent

GRS Viral DNA/RNA Purification Kit

GRS FullSample Purification Kit

Plasmid Purification

GRS Plasmid Purification Kit – Mini

GRS Plasmid Purification Kit – Transfection Grade

Enzymes

Proteinase K

RNase A

DNase I

Zymolyase

rSAP

Exonuclease I

Columns

Nucleic Acid Purification columns

26

27

27

28

28

29

30

31

32

32

33

33

34

34

35

14

Application/Kit PCR Purification Gel Extraction Column High-Throughput Enzymatic Dye Removal


Cut & Spin Gel Extraction Columns

Exo/SAP Go PCR Purification Kit

GRS Dye Terminator Removal Resincolumns not included

Application/Kit DNA RNA Plasmid Blood Animal Cells Tissue FFPE Bacteria Yeast/Fungus Plant Fluids/Cell Free DNA Card Swab Clean-Up Transfection Protein

GRS Genomic DNA Kit - Blood & Cultured Cells

GRS Genomic DNA Kit - Tissue

GRS Genomic DNA Kit - Plant

GRS Genomic DNA Kit - BroadRange

GRS Genomic DNA Kit - Bacteria

GRS Pure DNA Kit


GRS Total RNA Kit - Blood & Cultured Cells

GRS Total RNA Kit - Tissue

GRS Total RNA Kit - Plant

GRS Total RNA Kit - Bacteria

GRS Total RNA Kit - Yeast & Fungus


GRS Pure RNA Kit

GRS microRNA Kit(microRNA)

tripleXtractor Reagent



GRS Plasmid Purification Kit - Mini

GRS Plasmid Purification Kit - Transfection Grade

PCR Purification & Gel Extraction

DNA Purification

RNA Purification

DNA & RNA Purification

Plasmid Purification

Suitable Adaptable

Dye Terminator Removal

S A M P L E T Y P E S

Nucleic Acid Purification kit selection Guide

NA Purification / Selection Guide

Symon

discontinued 1


www.grisp.pt

15

Application/Kit PCR Purification Gel Extraction Column High-Throughput Enzymatic Dye Removal


Cut & Spin Gel Extraction Columns

Exo/SAP Go PCR Purification Kit

GRS Dye Terminator Removal Resincolumns not included

Application/Kit DNA RNA Plasmid Blood Animal Cells Tissue FFPE Bacteria Yeast/Fungus Plant Fluids/Cell Free DNA Card Swab Clean-Up Transfection Protein


GRS Genomic DNA Kit - Tissue




GRS Pure DNA Kit








GRS Pure RNA Kit

GRS microRNA Kit(microRNA)

tripleXtractor Reagent



GRS Plasmid Purification Kit - Mini

GRS Plasmid Purification Kit - Transfection Grade

S A M P L E T Y P E S

NA Purification / Selection Guide

16

The GRS PCR & Gel Band Purification Kit provides an effi-cient and fast method for the purification and/or concentra-tion of high quality DNA fragments (70bp to 15kb) from PCR reactions, enzymatic restriction digestion, or from agarose gels. Eluted DNA is suitable for all common downstream applications including PCR, enzymatic restriction diges-tion, cloning, library construction, Southern blot analysis, and DNA sequencing.

GRS PCR & Gel Band Purification KitSample:

Expected Yield:

Format:

Operation Time:

Elution Volume:

up to 100μL of PCR products or DNA fragments from enzymatic reactions up to 300mg of agarose gel typical recoveries up to 95% (PCR Clean-up) or up to 90% (Gel Extraction)spin column

10 minutes (PCR clean-up), 20 minutes (Gel Extraction)

20-50 μL

High pH Optimal pH

x √pH indicator to ensure optimal pH for DNA binding

Gel PCR Product Dissociation DNA Binding Wash Elution

or


GK01.0100 GRS PCR & Gel Band Purification Kit

100 preps/kit


3x 100 preps/kit


5x 100 preps/kit

NA Purification / PCR Purification & Gel Extration


www.grisp.pt

17


GK18.0500 Exo/SAP Go-PCR Purification Kit

500 rxn

GK18.2000 Exo/SAP Go-PCR Purification Kit

4x 500 rxn

The Exo/SAP Go - PCR purification kit is an enzymatic PCR Clean-Up kit, comprising Exonuclease I (Exo I) and recombinant Shrimp Alkaline Phos-phatase (rSAP) in an optimal molar ratio. Unused primers are hydrolyzed by Exo I, whilst rSAP dephosphorylates excess dNTPs.

Exo/SAP Go is designed to degrade dNTPs and primers in a single reac-tion tube in only 5 minutes. Both enzymes are subsequently inactivated by heating and DNA is ready for sequencing in 15 minutes. No further treatment is required and recovery is 100%, even for very short PCR products.

Exo/SAP Go - PCR Purification KitSample:

Expected Yield:

Format:

Operation Time:

5μL of PCR product

100% purification

enzymatic

15 minutes

high-throughput

100% product purification

fast (15 minutes)

economic

100% amplicon recovery

A

AA

A

AA

TTT

T

TT

TT

T

TT

C

C

CC

CC

G

GGG

GG

AA

A

AA

TT

T

TT

C

CC

CC

G

GG

GG

Unpurified PCR Product Purified PCR Product

15 minutes

Exo/SAP Go

1,1kb 400bp 80bp

Pre Post Pre Post Pre Post


18

Cut&Spin Gel Extraction ColumnsWith Cut&Spin columns in 5-10 minutes you have your DNA ready-to-go!Cut&Spin Gel Extraction columns are ready-to-use columns. Just cut out the DNA-band from the agarose gel, place it on the top of the column matrix, and centrifuge at 5000-6000g for 5-10 minutes at room temperature. The eluate containing the DNA can be used directly for subsequent applications such as cloning, RFLP, and sequencing.

A few restriction enzymes might be inhibited by EDTA present in TAE or TBE. If this is the case or if concentration is too low, DNA can be concentrated by common precipitation methods and dissolved in 10mM Tris-HCl or TE pH 8.0.

Advantage of GRiSP Cut&Spin Columns

✓ No need for special (low melting) agaroses.

✓ No melting or lysation procedures to dissolve agarose.

✓ No use of additional buffers or solutions.

✓ No use of vacuum manifolds or subsequent centrifugation procedures.

✓ No desalting procedures.

✓ Can be used for small sized DNA up to greater than 20 kbp.

✓ DNA directly usable for subsequent experiments such as cloning, RFLP, and sequencing

✓ Very fast (10 minutes)

✓ Simple protocol Gel Slice Cut & Spin Column Centrifugation DNA Collected in the buffer


GS131.0250 Cut&Spin Gel Extraction Columns

250 columns

GS131.0500 Cut&Spin Gel Extraction Columns

500 columns


GD05.0025 Xtractor Gel Band Cutter 25 units

Xtractor Gel Band CutterXtractor Gel Band Cutters are disposable tools that allow to excise DNA or RNA bands accurately from agarose gels without the risk of cross-contamination. For one-handed, uniform, and fast cutting, they provide an easy and safer alternative for razor blades or scalpels, as they do not scratch the glass of a transilluminator or cut into skin.

✓ No risk of cross contamination

✓ Fast & Accurate



www.grisp.pt

19


GD05.0025 Xtractor Gel Band Cutter 25 units


GK27.0100 GRS Dye TerminatorRemoval Resin

100g

GK27.0500 GRS Dye Terminator Removal Resin

500g

GRS Dye Terminator Removal ResinFor purification of samples prior to Sanger SequencingAfter DNA sequencing reactions, it is important to remove unincorpo-rated dyes and salts as they interfere with capillary electrophoresis resulting in errors and poor signal-to-noise ratios. GRS Dye Terminator Removal Resin is a matrix for size-exclusion chromatography based on a beaded composite material comprised of ultrapure cross-linked dextran. In gel-filtration, molecules are separated according to size with larger molecules passing through the column significantly faster than smaller molecules. The GRS Dye Terminator Removal Resin demonstrates chem-ical stability and low non-specific adsorption. Effects of Buffer and pH on resolution are not significant and dilution of purified macromolecules is only minimal (approximately 1,5-fold). The molecular weight cut-off for these beads is 20 base pairs, making the GRS Dye Terminator Removal Resin an excellent choice for the clean-up of unincorporated dye termi-nators and salts of sequencing reactions before capillary electrophoretic injection.

GRS Dye Terminator Removal Resin is presented as a powder that can be hydrated in aqueous media containing up to 20% of ethanol.

One gram of powder is sufficient for the preparation of approximately 10 ml of slurry.

NA Purification / Dye Terminator Clean-Up

20

The GRS Genomic DNA Kit – Tissue - provides an efficient and fast method for the purification and/or concentration of high quality total DNA (including genomic DNA, mitochondrial DNA and viral DNA) from a variety of samples, including tailsnips, liver, kidney, brain, adipose tissue, earpunches, insects, and FFPE. Eluted purified DNA (approximately 20-30kb) is suitable for PCR, and other enzymatic reactions.Proteinase K and RNase A included

GRS Genomic DNA Kit - TissueSample:

Expected Yield:

Format:

Operation Time:

Elution Volume:

up to 30mg of tissue or 25mg of paraffin-embedded tissue.up to 50µg DNA

spin column

within 30 minutes (60 min for FFPE)

50-200 μL

Blood RBC Lysis Pellet Cell Lysis DNA Binding Wash Elution

Homogenization Cell Lysis DNA Binding Wash Elution

The GRS Genomic DNA Kit – Blood & Cultured Cells - provides an efficient and fast method for the purif ication and/or concentration of high quality total DNA (including genomic DNA, mitochondrial DNA and viral DNA) from a variety of samples, including whole and frozen blood, buffy coat and cultured animal cells. Eluted purified DNA (approximately 20-30kb) is suitable for PCR, and other enzymatic reactions.Proteinase K and RNase A included

GRS Genomic DNA Kit - Blood & Cultured CellsSample:

Expected Yield:

Format:

Operation Time:

Elution Volume:

up to 300μL of whole fresh blood, up to 5x106 cultured animal cells2-3µg (300 μL blood) or 25-30µg (1x106 293T cells) of total DNA

spin column

within 20 minutes

25-100μL


GK02.0100 GRS Genomic DNA Kit Blood & Cultured Cells

100 preps/kit


GK03.0100 GRS Genomic DNA Kit Tissue

100 preps/kit

NA Purification / gDNA Purification


www.grisp.pt

21



GK03.0100 GRS Genomic DNA Kit Tissue

100 preps/kit


GK04.0100 GRS Genomic DNA Kit Plant

100 preps/kit


GK07.0100 GRS Genomic DNA Kit Bacteria

100 preps/kit

The GRS Genomic DNA Kit - Plant - provides an efficient and fast method for the purification and/or concentration of high quality total DNA (including genomic DNA, mitochon-drial DNA, and chloroplast DNA) from plant tissue and cells. Eluted purified DNA is suitable and ready-to-use for PCR, real-time PCR, Southern Blotting, and RFLP.

RNase A included

The GRS Genomic DNA Kit – Bacteria – provides an efficient and fast method for the purification of high quality genomic (and viral) DNA from Gram(+) positive and Gram(-) negative bacteria, suitable for al l common downstream applica-tions such as PCR, enzymatic restriction digestion, cloning, Southern blot analysis, etc.

Lysozyme and RNase A included



Sample:

Expected Yield:

Format:

Operation Time:

Elution Volume:

Sample:

Expected Yield:

Format:

Operation Time:

Elution Volume:

up to 100mg of fresh plant tissue, up to 25 mg of dry plant tissue3-5µg total DNA from 100mg Arabidopsis thaliana young leaf samples; 20-25µg total DNA from 100mg Nicotiana tabacum young leaf samplesspin column

within 60 minutes

100μL

0,5-2,0 ml of bacterial culture with up to 1x109 cells (Gram (+) or Gram (-))25-30 μg (1x109 of Escherichia coli); 10-15 μg (1x109 of Bacillus subtilis)spin column less than 1 hour 30-200 μL

Sample Cell Lysis Filter Eluate DNA Binding Wash Elution

Pellet Cell Lysis DNA Binding Wash Elution


22

The GRS Genomic DNA Kit – BroadRange - provides an efficient and fast method for the purification of high quality total DNA (including genomic DNA, mitochondrial DNA and viral DNA) from a broad range of samples, including whole and frozen blood, serum, plasma, buffy coat, amniotic fluid, buccal swab, hair, tissue, rodent tails, ear punches, FFPE, cultured cells and insect cells. Eluted purified DNA (approximately 20-30kb) is suitable for PCR, and other enzymatic reactions Proteinase K and RNase A included

The GRS Pure DNA Kit provides an efficient and fast method for the purification and/or concentration of high quality DNA (50bp to 30kb) from samples containing partial purified DNA (genomic DNA, mitochondrial DNA, PCR products, etc) obtained via other DNA isolation methods. The purified DNA is suitable for all common downstream applications including PCR, RFLP, cloning, library construction, Southern blot analysis, and DNA sequencing. Purify and/or concentrate partially purified DNA samples


GRS Pure DNA Kit

Sample:

Expected Yield:

Format:Operation Time:Elution Volume:

Sample:Expected Yield:Format:Operation Time:Elution Volume:

up to 200μL of whole blood (fresh or frozen), serum, plasma, buffy coat or other body fluids, up to 25 mg of tissue, up to 25mg of paraffin-embedded tissue (FFPE), up to 15ml of amniotic fluid, buccal swab, hair, cultured cells and insect cells.5 µg DNA (in case of whole fresh blood) spin columndepending on the type of sample50-200μL

up to 100μL of DNA products up to 90% spin column20 minutes20-50 μL

DNA Sample DNA Binding Wash Elution

Sample Cell Lysis DNA Binding Wash Elution


GK06.0100 GRS Genomic DNA Kit BroadRange

100 preps/kit


GK15.0100 GRS Pure DNA Kit 100 preps/kit



www.grisp.pt

23

The GRS Genomic DNA Kit – Card provides an easy and efficient method for the purification of high quality genomic DNA from dried blot spots (on Whatman® FTA® Cards). Eluted purified DNA (approximately 20-30kb) is suitable for PCR, real-time PCR, and other enzymatic reactions.

The GRS Genomic DNA Kit – Card uses proteinase K and chaotropic salts to lyse cells and to denature proteins. Carrier RNA is included to minimize DNA loss to plasticware. The buffer system is optimized to allow selective binding of DNA to the glass fiber matrix of the spin column. Contaminants such as proteins, divalent cations, unincorpora-ted nucleotides, and enzyme inhibitors are completely removed using a Wash Buffer (containing ethanol) in a simple centrifugation step. The purified genomic DNA is subsequently eluted by a low salt Elution Buffer or TE or water. The entire procedure can be completed within an hour without phenol/chloroform extraction or alcohol precipitation, with typical DNA yields of 300ng from a 6mm dried blot spot.

Sample:Expected Yield:

Format:

Operation Time:

Elution Volume:

dried blood spots (on Whatman® FTA® Card)typically more than 300ng of pure genomic DNA from 6mm dried blood spotsspin column

approximately 1 hour.

30-100 μL


GK25.0100 GRS Genomic DNA Kit Card

100 preps/kit


DNA Card Cell Lysis DNA Binding Wash Elution


24

The GRS Total RNA Kit – Blood & Cultured Cells - provides an efficient and fast method for the purification of high quality total RNA (including mRNA, tRNA and rRNA) from fresh whole blood and cultured animal cells. Eluted purified RNA is suitable for RT-PCR, Northern Blotting, mRNA selection, cDNA synthesis, and primer extension.DNase I included Individually packed columns

The GRS Total RNA Kit - Tissue - provides an efficient and fast method for the purification and/or concentration of high quality total RNA (including mRNA, tRNA and rRNA) from a variety of animal and paraffin-embedded tissues. Option-al DNase treatment can be included in the protocol to remove residual DNA. Eluted purified RNA is suitable for RT-PCR, Nor thern B lot t ing , mRNA se lect ion , cDNA synthes is , and primer extension.Proteinase K and DNase I included Individually packed columns



Sample:

Expected Yield:

Format:

Operation Time:

Elution Volume:

Sample:

Expected Yield:

Format:

Operation Time:

Elution Volume:

up to 300μL of whole fresh blood, up to 5x106 cultured animal cells2-3µg (300 μL blood) or 25-30µg (1x106 293T cells) of total RNAspin column

within 20 minutes

25-100μL

up to 25mg of tissue up to 25 mg of paraffin-embedded tissue5-30µg total RNA

spin column

within 60 minutes

50μL


GK08.0100 GRS Total RNA Kit Blood & Cultured Cells

100 preps/kit


GK09.0100 GRS Total RNA Kit Tissue

100 preps/kit

RBC Lysis Pellet Cell Lysis RNA Binding Wash Elution

Homogenization Cell Lysis RNA Binding Wash Elution

NA Purification / RNA Purification


www.grisp.pt

25

The GRS Total RNA Kit - Plant - provides an efficient and fast method for the purification and/or concentration of high quality total RNA (including mRNA, tRNA and rRNA) from plant tissue and cells. Optional DNase treatment can be included in the protocol to remove residual DNA. Eluted purified RNA is suitable for RT-PCR, One-step qRT-PCR, Northern Blotting, mRNA selection, cDNA synthesis, and primer extension.

DNase I included Individually packed columns

The GRS Total RNA Kit – Bacteria - provides an efficient and fast method for the purification of high quality total RNA (including mRNA, tRNA and rRNA) from Gram(-) negative and Gram(+) positive bacteria. Eluted purified RNA is suitable for RT-PCR, Northern Blotting, mRNA selection, cDNA synthesis, and primer extension.Lysozyme and DNase I included Individually packed columns



Sample:

Expected Yield:

Format:

Operation Time:

Elution Volume:

Sample:

Expected Yield:

Format:

Operation Time:

Elution Volume:

up to 100mg of fresh plant tissue, up to 25 mg of dry plant tissue5-30µg total RNA from young leaf samples

spin column

within 30 minutes

50μL

up 1x109 bacterial cells [Gram(-) negative or Gram(+) positive bacteria].up to 60µg total RNA (40-45 µg in case of 1x109 Escherichia coli 45-55 µg in case of 1x109 Bacillus subtilis)spin column

within 20 minutes

50-100μL


GK10.0100 GRS Total RNA Kit Plant

100 preps/kit


GK16.0100 GRS Total RNA Kit Bacteria

100 preps/kit

Sample Cell Lysis RNA Binding Wash Elution

Pellet Cell Lysis RNA Binding Wash Elution


26

The GRS Total RNA Kit – Yeast & Fungus - provides an effi-cient and fast method for the purification of high quality total RNA (including mRNA, tRNA and rRNA) from yeast and a wide variety of other fungus species. Eluted purified RNA is suit-able for RT-PCR, Northern Blotting, mRNA selection, cDNA synthesis, and primer extension.DNase I included Individually packed columns

The GRS Pure RNA Kit provides an efficient and fast method for the puri-fication and or concentration of high quality RNA from samples contain-ing partial purified RNA obtained via other RNA isolation methods. Contaminants (e.g. genomic DNA or residual phenol) are effectively removed in less than 15 minutes , with a typical RNA recovery of70- 80%. Purified RNA is suitable for use in Northern Blotting and RT-PCR. High Quality RNA (A260/A280 of 1.9-2.0) Individually packed columns


GRS Pure RNA Kit

Sample:

Expected Yield:

Format:

Operation Time:

Elution Volume:

Sample:

Expected Yield:

Format:

Operation Time:

Elution Volume:

up to 5x107 yeast or other fungus cells.

up to 30µg total RNA

spin column

20-30 minutes from spheroplast; 1h15-1h30 from whole cells50-100μL

up to 100μL of RNA products

up to 50µg

spin column

15 minutes

20-50 μL


GK17.0100 GRS Total RNA Kit Yeast & Fungus

100 preps/kit


GK15.0100 GRS Pure RNA Kit 100 preps/kit

Pellet Cell Lysis RNA Binding Wash Elution

RNA Sample RNA Binding Wash Elution



www.grisp.pt

27

Sample:

Expected Yield:

Format: Operation Time:

Elution Volume:

Up to 200 μL of body fluids (whole blood, buffy coat, serum, etc) up to 5x106 cultured animal yeast or plant cells, 50-100mg of tissue, up to 1x109 bacterial cells.total RNA (e.g. up to 3 µg from 200 μL of whole human blood, up to 20 µg from 1x109 cells of Escherichia colispin column

within 15minutes 20-50μL

The tripleXtractor directRNA kit combines the strong lysis capability of the monophasic phenol/guanidine thiocyanate solution “tripleXtractor Direct” with a spin column system for convenient, rapid and cost effec-tive purification of ultrapure RNA without the need of chloroform phase separation and isopropanol precipitation.

The purified total RNA is free of DNA and proteins, and can be used for all common downstream applications such as cDNA Library construc-tion, Northern Blotting, RT-PCR, in vitro translation, Nuclease Protection Assays, etc.DNase I included Individually packed columns

Standard protocols for the isolation of total RNA or mRNA are not optimized for the purification of small RNA molecules, resulting in the loss of consi derable amounts of micro RNAs (miRNAs) and other small cellular RNAs. Moreover, accurate analysis of expression of miRNAs by qPCR or by microarray analysis requires the removal of the larger RNA molecules. The GRS microRNA Kit provides an efficient and fast method for the purification of high quality miRNAs (and other small cellular RNAs), with minimal contamination from large RNA molecules and genomic DNA, from a variety of tissues and cultured cells. Individually packed columns


GRS microRNA Purification Kit


GK23.0100 tripleXtractor direct RNA Kit

100 preps/kit


GK11.0050 GRS microRNA Purification Kit

50 preps/kit

Lysis RNA Binding Wash Elution

Sample:

Expected Yield:

Format:

Operation Time:

Elution Volume:

up to 100mg of tissue, up to 1x106 cultured

miRNA (and other small cellular RNAs) spin column

30 minutes

50 μL

Cell Lysis RNA Binding miRNA Binding Wash Elution

Direct purification from tripleXtractor without phase separation


28

TripleXtractor is a cost-effective, ready-to-use reagent for the isolation of high-quality total RNA from cells and tissues. DNA and/or proteins can be subsequently recovered by sequential precipitation. TripleXtractor can be used with a wide variety of samples, including blood, buffy coat, cultured cells, animal and plant tissues, bacteria and yeast.

The purified total RNA is free of DNA and proteins and can be used for all common downstream applications such as cDNA Library construc-tion, Northern Blotting, RT-PCR, in vitro translation, nuclease protection assays, etc.Efficient and economic

The GRS Viral DNA/RNA Purification Kit provides an effi-cient and fast method for the purification of high quality viral DNA and RNA from cell-free media (e.g. from serum, plasma, body fluids, and the supernatant from viral infected cell cultures). Eluted purified Nucleic Acid is suitable for all common applications, including PCR, real-time PCR, RT-PCR, One-step qRT-PCR, and DNA Sequencing. This kit is recommended for parallel purifica-tion of viral DNA (including CMV and HBV) and viral RNA (including HIV, HTLV, and HCV). The detection limit depends on the type of virus and on the sensitivity of individual PCR or RT-PCR protocols.

tripleXtractorReagent for RNA Isolation


Sample:

Expected Yield:

Operation Time: Elution Volume:

Sample:

Expected Yield:

Format: Operation Time:

Elution Volume:

whole blood, buffy coat, cultured animal cells or tissue, cultured plant cells or tissue, cultured bacterial cells, cultured yeast cells. total RNA (optional: also DNA and/or proteins)

within 60 minutes 10-50 μL

200µl of cell-free medium (serum, plasma. body fluid, supernatant of viral infected cells)

viral DNA and RNA

spin column within 40 minutes 50μl


GB23.0100 tripleXtractor Reagent for RNA Isolation

100 mL


GK12.0050 GRS Viral DNA/RNA Purification Kit

50 preps/kit

Lysis Phase Separation Precipitation Wash RNA Resuspension

DNA, RNA, and proteins, by phase separation

Sample Cell Lysis Binding Wash Elution

NA Purification / DNA & RNA Purification


www.grisp.pt

29

The GRS FullSample Purification Kit provides an efficient and fast method for the simultaneous purification of genomic DNA, total RNA (including miRNA), and total protein from whole blood and other biolog-ical fluids, animal tissues, and cultured cells. DNA is bound to a DNA binding spin column and the flow-through is subsequently passed through a RNA binding spin column. The proteins in the flow-through can then be precipitated with acetone. Contaminants are washed away and DNA is eluted with a low salt buffer and RNA is eluted with RNase-free water. The entire procedure can be completed in less than half an hour without phenol/chloroform or alcohol precipitation. Eluted gDNA (20-30kb) is suitable for most common downstream applications, includ-ing PCR, whereas purified total RNA (including microRNA) is ready to use for RT-PCR, primer extension, and Northern Blotting. After protein precipitation, the pellet is washed with ice-cold ethanol and resuspend-ed in an appropriate volume of desired buffer compatible with the down-stream application of choice.

GRS FullSample Purification KitSample:

Expected Yield: Format:

Operation Time: Elution Volume:

up to 200μL of biological fluids , 500 μL whole blood), 25mg of animal tissue, or 5x106 cultured animal cellstypically up to 9µg of genomic DNA , 20µg of total RNA, and 120µg of protein (from 1,5x106 HeLa cells).spin column

25 minutes for DNA & RNA, approximately 1 hour for protein precipitation.50-200 μL for DNA, 25-50 μL for RNA, protein resuspension in desired volume.


GK26.0050 GRS FullSample Purification Kit

50 preps/kit

Fig 1/ Total RNA and Genomic DNA were extracted from 1,5x106 HeLa cells using the GRS FullSample Purification Kit, and 10μL aliquots of purified RNA (total 50μL) and of purified DNA (total 200μL) were subse-quently analyzed by electrophoresis on a 1% agarose gel. Yields were determined by spectrophotometry: a total of 20,9 µg of total RNA was purified with an A260/A280 of 2,09 and A260/A230 of 12,03 as well as 9,3µg of genomic DNA with an A260/280 of 1,88 and A260/A230 of 2,18.

Fig 2/ Total protein was extracted from 1,5x106 HeLa cells using the GRS FullSam-ple Purification Kit, and a 20μL aliquot of purified protein (total 200μL) was subse-quently analyzed by SDS-PAGE, stained with Coomassie Brilliant Blue. Yield was deter-mined by the Bradford protein assay: 120µg (at a concentration of 0,6µg/μL).

68

53

41

32

23

14

10

ICE

Sample Preparation And Cell Lysis

DNA Wash DNA Elution

DNA/RNA Separation: DNA Binding & RNA

Flow-trough

RNA Binding and Protein Flow-trough

RNA Wash RNA Elution

Protein Precipitation

Protein Resuspension

NA Purification / DNA & RNA Purification

30

The GRS Plasmid Purification Kit provides an efficient and fast method for the purification of high quality plasmid DNA from 1-6 ml of cultured bacterial cells (miniprep). Eluted DNA is suitable for all common down-stream applications including PCR, enzymatic restriction digestion, cloning and DNA sequencing.

GRS Plasmid Purification Kit - MiniSample:

Expected Yield: Format: Operation Time: Elution Volume:

1,5 ml of cultured bacterial cells (up to 6 ml)

up to 50 µg of plasmid DNA spin column within 20 minutes 30-100 μL


GK13.0100 GRS Plasmid Purification Kit - MINI

100 preps/kit


3x 100 preps/kit


5x 100 preps/kit

Includes Blue Lysis Buffer for easy visualization of lysis and neutralization

Incomplete Lysis

Incomplete Neutralization

Complete Lysis

Complete Neutralization

x

x

√

√

Cell Harvesting Cell Lysis DNA Binding Wash Elution

NA Purification / Plasmid Purification


www.grisp.pt

31

The GRS Plasmid Purification Kit – Transfection Grade provides an easy and efficient method for the purification of high quality plasmid DNA from cultured bacterial cells. Eluted DNA is free of RNA and endotox-ins and is suitable for transfection and other downstream applications including PCR, enzymatic restriction digestion, cloning, in-vivo transcrip-tion, and DNA sequencing.

The GRS Plasmid Purification Kit – Transfection Grade is based on a modified Alkaline Lysis method used to obtain minimal genomic DNA contaminants. RNase A treatment reduces contamination with RNA. The buffer system is optimized to allow binding of plasmid DNA in the presence of chaotropic salts to the anion-exchange column. Contam-inants are removed using a Wash Buffer. The purified plasmid DNA is subsequently eluted using a high salt buffer and then isopropanol-pre-cipitated for desalting. The entire procedure can be completed in within 90 minutes without phenol/chloroform extraction or the need of an ultra-centrifuge with a typical DNA yields of 1000µg of transfection grade plasmid..

GRS Plasmid Purification Kit – Transfection GradeSample:

Expected Yield: Endotoxin: Format: Operation Time: Elution Volume:

50-400 ml of cultured bacterial cells harbouring target plasmidtypically 1000 µg of transfection grade plasmid DNA <0.1 EU/µg DNA gravity –flow IEX column within 90 min.

0,5 ml – 2,0 ml


GK73.0002 GRS Plasmid Purification Kit - Transfection Grade

2 preps/kit


10 preps/kit


25 preps/kit

Cell Culture Alkaline Lysis Equilibration/Binding Wash/Elution DNA Precipitation

Fast ProcedurePre-filter includedTransfection Grade DNA

Very low endotoxin levelsVery high yields

NA Purification / Plasmid Purification

Symon

sorry

32

Proteinase K(lyophilized, -30KU/mg)

Proteinase K is a broad-spectrum serine protease that cleaves peptide bonds at the carboxylic sides of aliphatic, aromatic, and hydrophobic amino acids.

Features

✓ Recombinant protein purified from Pichia pastoris harbouring the gene encoding endolytic protease (proteinase K, 28,9kDa mono-mer) from Tritirachium album

✓ Lyophilized powder with Specific Activity of ~30 Kunitz Units per mg

Applications

✓ Isolation of genomic DNA from cultured cells and tissues

✓ Removal of DNases and RNases during DNA and/or RNA purification

✓ Determination of enzyme locations.


GE010.0100 Proteinase K (lyophi-lized, ~30KU/mg)

100mg


GE011.0100 RNase A (DNase and Proteinase free, lyo-philized, >50KU/mg)

100mg

RNase A(DNase and Proteinase free, lyophilized, >50KU/mg)

RNase A (Ribonuclease A) is a pancreatic endoribonuclease (13.7 kDa monomer) that specifically cleaves single-stranded RNA at the 3’ end of pyrimidine residues. The phosphodiester bond between the 5’-ribose of one nucleotide and the phosphate group of the 3’-ribose of an adjacent pyrimidine nucleotide is broken and the resulting 2’, 3’-cyclic phosphate is hydrolyzed to the corresponding 3’-nucleoside phosphate.

Features

✓ Purified from Bovine Pancreas.

✓ Lyophylized powder with specific activity > 100Kunitz units per mg (>5000 Worthington units/mg)

✓ DNase-free (no need to heat the RNase before use)

✓ Proteinase-free

Applications

✓ Isolation of genomic and plasmidic DNA from cultured cells and tissues

✓ Removal of RNA from recombinant protein preparations

✓ Ribonuclease protection assays

✓ Mapping of mutations in DNA or RNA by mismatch cleavage. RNase A cleaves the RNA in RNA:DNA hybrids at sites of single base mismatches, and the cleavage products can be analyzed.

NA Purification / Enzymes


www.grisp.pt

33


GE012.0100 DNase I (lyophilized, RNase-free, >2000KU/mg, with storage and reaction buffers)

100mg


GE013.0001 Zymolyase® - 20T 1g

Features

✓ Recominant enzyme, purified from E.coli harbouring an MBP fusion clone of Bovine Pancreatic DNase I (29kDa monomer)

✓ Lyophilized powder with specific activity: >2000 Kunitz units per mg.

✓ RNase-free

✓ Supplied with 10 ml of 10x reaction buffer and with 1 ml of storage buffer; Reaction buffer (10x) is 100 mM Tris-HCl (pH 7.5 at 25°C), 25 mM MgCl2, 1 mM CaCl2. Storage buffer is 10 mM Tris-HCl (pH 7.5), 10 mM CaCl2 10 mM MgCl2, 50% (v/v) glycerol

Applications

✓ Removal of residual genomic DNA from RNA samples

✓ Degradation of DNA template in transcription reactions

✓ DNase I footprinting

✓ Nick Translation

Zymolyase® - 20T from GRiSP is prepared from Arthrobacter luteus, and is supplied as an ammonium sulfate precipitate of a complex of enzymes. The strong lytic activity against living yeast cell walls is mainly due to the activity of ß-1,3-glucan laminaripentaohydrolase. This enzyme hydrolyz-es linear glucose polymers with β-1,3-linkages and releases specifically laminaripentaose as the main and minimum product unit, allowing for the production of protoplasts or spheroplasts of various yeast strains.

Lytic activity varies depending on yeast strain, growth stage of yeast, or cultural conditions. Under the conditions mentioned below, the lytic activity of Zymolyase® - 20T is 20.000 U/g.

Applications

✓ Preparation of spheroplasts or protoplasts from variety of yeast strains

NOTE: Zymolyase® is a registered trademark of Kirin Holdings Company, Limited.

DNase I(lyophilized, RNase-free, >2000KU/mg, with storage and reaction buffers)

Zymolyase® - 20T

DNase I (Deoxyribonuclease I) is an endonuclease that cleaves DNA pre ferentially at phosphodiester linkages adjacent to a pyrimidine nucleotide, to release di-, tri-, and oligonucleotide products (on average producing tetranucleotides) with 5’-phosphorylated and 3’-hydroxylat-ed ends. DNase I acts on single-stranded DNA, double-stranded DNA, RNA-DNA hybrids, and chromatin.


Symon

PRODUCT CHANGE

34

Recombinant Shrimp Alkaline Phosphatase (rSAP) is a heat-labile multi-purpose alkaline phosphatase that catalyzes the dephosphory-lation of DNA, RNA, nucleotides, and proteins. With respect to other alkaline phosphatases, rSAP is the enzyme of choice as it, unlike Calf Intestinal Alkaline Phosphatase, can be heat-inactivated, and in compar-ison to Antartic Phosphatase, it has a much higher activity in most of the common enzyme reaction buffers, including PCR reaction mixtures, and does not require buffer modifications for activity. In comparison with native SAP, recombinant SAP is far more stable at room temperature and moreover available at higher concentrations.

Exonuclease I (Exo I) is an exonuclease that hydrolyzes single-stranded DNA, one nucleotide at a time from the end (exo) in the 3´>5´ direction. It releases 5´-mononucleotides one after another and leaves the terminal 5´-dinucleotide intact. This enzyme, product of the sbcB gene of E.coli, does not cleave DNA strands without terminal 3´-OH groups, since these are blocked by phosphoryl or acetyl groups. It does not degrade double-strand-ed DNA. For activity, Exo I requires magnesium (optimal concentration is 10mM) and since Exo I is tolerant to a wide variety of buffer conditions, it can be added directly to most molecular biology buffers containing magnesium (>1mM), including PCR reaction buffers.

Shrimp Alkaline Phosphatase (rSAP)

Exonuclease I


GE015.0001 Shrimp Alkaline Phosphatase (1U/μL)

1000 U


GE014.0001 Exonuclease I (20U/μL) 20000 U

Applications

✓ Removal of dNTPs from PCR reaction prior to DNA sequencing or labelling (typically in combination with exonclease I (Exo I))

✓ Dephosphorylation of DNA prior to end-labelling (using T4 Polynu-cleotide Kinase)

✓ Dephosphorylation of vectors (plasmids) during cloning, in order to prevent recircularization during ligation reaction (using T4 DNA Ligase).

Features

✓ Recombinant protein purified from Pichia pastoris harbouring the gene encoding shrimp alkaline phospatase form Panda-lus borealis (54kDa)

✓ 1ml solution with Specific Activity of 1 unit per microliter (20.000 units in total).

✓ Free of RNase, exonucleases, and endo-nuclease activities

Applications

✓ Removal of primers from PCR reaction prior to DNA sequencing or labelling (typically in combination with rSAP)

✓ Removal of linear DNA molecules form plasmid preparation (typi-cally in combination with lambda exonuclease)

✓ Removal of single-stranded DNA containing a 3´-hydroxyl group from heterogeneous mixtures of nucleic acids.

✓ Assay for the presence of single-stranded DNA regions

Features

✓ Recombinant protein purified from Escheri-chia coli harbouring the gene encoding exonuclease I (55kDa)

✓ 1ml solution with Specific Activity of 20 units per ul (20.000 units in total), supplied with 1,5ml of 10x reaction buffer (670mM Glycine-KOH pH9,5, 67mM MgCl2, and 100mM beta-mercaptoethanol)

✓ Free of RNase and endonuclease activities



www.grisp.pt

35

For your convenience, GRiSP also supplies Nucleic Acid Purification columns separately:


GKC.PG50 PCR Purification & Gel Extraction Columns 50 Units

GKC.GC50 Genomic DNA mini Columns 50 Units

GKC.GF50 Genomic DNA filter Columns 50 Units

GKC.RC50 RNA mini Columns (individually packed) 50 Units

GKC.RF50 RNA filter Columns 50 Units

GKC.VC50 Viral mini Columns (individually packed) 50 Units

GKC.PN50 Plasmid Purification mini Columns 50 Units

NA Purification / Columns

DNA Amplification

PCR Enzymes & Mastermixes

GRS Taq DNA polymerase

Xpert Taq DNA Polymerase

Xpert Fast DNA Polymerase

Xpert Hotstart DNA Polymerase

Xpert HighFidelity DNA Polymerase

qPCR

Xpert Fast SYBR

Xpert Fast Probe

Nucleotides

dNTP mix

dNTP Set

39

40

40

41

41

42

43

44

44

03Water

GRS PCR Grade Water

PCR Plastics

PCR tubes (flat cap)

PCR strips (attached flat caps)

qPCR plates

qPCR seals

44

45

45

45

45

38

The product range of Xpert DNA polymerases is comprised of a series of DNA polymerases with specific characteristics, designed to perform with excellent results for your most demanding applications.

As choosing the right enzyme for each particular application is an essen-tial part of the process, GRiSP presents below some guidelines on how to choose the right enzyme for your needs:

Main Applications Specificity Fidelity vs Taq Extension Rate Hotstart A-overhang Product Size

GRS Taq Routine PCR + 1X 1 kb/min - yes up to 5Kb

Xpert Taq Improved Routine PCRTA CloningGenotyping

++ 1X 1 kb/min - yes up to 10Kb

Xpert Fast Fast Routine PCRTA Cloning

++ 1X 4 to 6 kb/min - yes up to 3Kb

Xpert Hotstart GC/AT - richComplex templatesLong PCRMultiplex PCR

+++ 18X 1 kb/min yes yes up to 15Kb

Xpert HighFidelity High-Fidelity PCR Blunt-end CloningSite-directed Mutagenesis

+++ 50X 2 kb/min - no (blunt) up to 10Kb

Symon

PRODUCT CHANGE


www.grisp.pt

39

GRS Taq DNA Polymerase ✓ For Routine amplifications

✓ Amplification of targets up to 5kb

✓ Incorporates modified nucleotides

GRS Taq DNA polymerase is a recombinant thermostable enzyme (94kDa monomer) isolated from an E.coli strain harbouring the DNA polymerase gene from Thermus aquaticus YT-1. This enzyme has iden-tical characteristics as native Taq regarding activity, specificity, thermo-stability, and performance in PCR. GRS Taq possesses 5´→3´ polymerase activity but shows no 3´→5´ exonuclease activity (thus no proofreading). GRS Taq is suitable for amplifying DNA targets up to 5kb and produc-es 3´ A-overhangs allowing TA-cloning. The enzyme is available at a concentration of 5U/μL supplied with separated buffer as well as in a convenient mastermix.


GE01.0500 GRS Taq DNA polymerase 500U

GE01.5500 GRS Taq DNA polymerase 5x 500U

GE02.0100 GRS Taq 2x Mastermix (100rxn 25uL)

1,25mL

GE02.5100 GRS Taq 2x Mastermix (500rxn 25uL)

5x 1,25mL

GRS Taq Xpert Taq Xpert Fast Xpert Hotstart Xpert HighFidelity

Main Applications Routine PCR Improved Routine PCRGenotypingTA Cloning

Fast Routine PCRTA Cloning

GC/AT-rich PCRComplex tem-platesLong PCRMultiplex PCR

High-Fidelity PCRBlunt-end CloningSite-directed Mutagenesis

Increased Yield Xpert Taq

Fast Cycling Xpert Fast

High Specificity Xpert Hotstart

Long amplifications Xpert Hotstart; Xpert HighFidelity

Demanding samples Xpert Hotstart; Xpert HighFidelity

Low error-rate Xpert HighFidelity

Comparing with GRS Taq DNA polymerase, all the DNA polymerases included in the Xpert product range allow for an improved performance, specially designed to overcome specific needs:

Choose the right option for your application:

DNA Amplification / PCR Enzymes & Mastermixes

40

Xpert Taq DNA Polymerase ✓ High yield and sensitivity

✓ Robust amplification

✓ Amplification of targets up to 10Kb

Xpert Taq DNA Polymerase is a DNA polymerase with enhanced perfor-mance, when compared to regular Taq, and is optimized for demanding routine amplifications. The robustness of Xpert Taq DNA polymerase, as well as its high sensitivity and yield, even for targets of 10kb, makes this the ideal enzyme for daily amplifications with improved results.

Cycling conditions for Xpert Taq DNA polymerase are similar to those for conventional Taq DNA polymerase, making is easy to convert a PCR from regular Taq to Xpert Taq, thus improving the amplification with minimal effort.

PCR products generated with Xpert Taq DNA polymerase contain 3’-A overhangs, and can thus be cloned into TA cloning vectors.

Xpert Fast DNA Polymerase ✓ High yields under fast PCR conditions

✓ Buffer including dNTPs


Xpert Fast DNA polymerase is a robust enzyme, ideal for daily applica-tions like genotyping and screening, amplifying with extreme speed, yield and consistency.

Xpert Fast DNA polymerase has 5’-3’ exonuclease activity, but no 3’-5’ exonuclease (proofreading) activity, thus the PCR products generated with this enzyme are A-tailed, and can thus be cloned into TA cloning vectors.

The extreme speed of Xpert Fast DNA polymerase allows the use of an extension rate of 4-6 kb/min, making this the ideal choice for consistent results in routine PCR amplifications.

The supplied buffer includes not only MgCl2, but also 5mM dNTPs, enhancers and stabilizers, optimized to increased PCR success rates.


GE03.0500 Xpert Taq DNA polymer-ase

500U

GE04.0100 Xpert Taq 2x Mastermix (100rxn 25uL)

1.25mL


GE05.0500 Xpert Fast DNA poly-merase

500U

Components

Kit Mastermix

Xpert Taq DNA polymer-ase (5U/uL)

2X Xpert Taq Mastermix

10X Xpert Taq reaction buffer

DMSO

DMSO MgCl2 (25mM)

MgCl2 (25mM)

Components

Xpert Taq DNA polymerase (5U/uL)

5X Xpert Fast reaction buffer (dNTPs included)



www.grisp.pt

41

Xpert Hotstart DNA Polymerase ✓ Amplification of complex targets

✓ Reduced non-specific amplification

✓ Multiplex PCR


Xpert Hotstart DNA Polymerase is a DNA polymerase combined with two proprietary proteins that bind to DNA, efficiently neutralizing the DNA polymerase activity at room temperature. At this temperature, one protein binds to double stranded DNA, while the other binds to primers, making this the most effective blocking method for hotstart. Xpert Hotstart DNA polymerase is thus the ideal choice for the amplifi-cation of complex targets, multiplex PCR, and all other situations when reduced non-specific amplification and reduced primer-dimer formation is essential.

Additionally, Xpert Hotstart DNA polymerase is able to efficiently amplify DNA fragments up to 15kb, making this an excellent option, especially when the size of the target is an issue.

PCR products generated with Xpert Hotstart DNA polymerase are A-tailed, and can thus be cloned into TA cloning vectors.

Xpert HighFidelity DNA Polymerase ✓ Extreme Fidelity (50x higher than Taq)

✓ Buffer including dNTPs

✓ Specific and fast amplification from complex templates

✓ Amplification of targets up to 10Kb

Xpert HighFidelity DNA polymerase is a robust enzyme with enhanced DNA binding, resulting in improved processivity, yield, and extremely low error-rate, ideal for applications such as high-fidelity PCR, site-di-rected mutagenesis, crude sample PCR, blunt-end cloning, among others where robustness and proof-reading are important.

The Xpert HighFidelity DNA polymerase has an error rate of approxi-mately 1 error per 4.5 x 107 nucleotides incorporated, which is 50x lower than Taq DNA polymerase.

The velocity of Xpert HighFidelity DNA polymerase allows the use of an extension rate of 2kb/min, making this also an ideal choice for consis-tent results in fast PCR amplifications.

The supplied buffer includes not only MgCl2, but also 5mM dNTPs, enhancers and stabilizers, optimized to increased PCR success rates.


GE06.0500 Xpert Hotstart DNA poly-merase

500U


GE07.0250 Xpert HighFidelity DNA polymerase

250U

Components

Xpert Hotstart DNA polymerase (2.5U/μL)

10X Xpert Hotstart reaction buffer

10X GC enhancer

Components

Xpert HighFidelity DNA polymerase (2U/μL)

5X Xpert HighFidelity reaction buffer (dNTPs included)


Symon

PRODUCT CHANGE

42

Xpert Fast SYBR ✓ Excellent signal with low PCR inhibition

✓ Early Ct values - Rapid extension rate

✓ Extreme sensitivity – increased limit of detection

✓ Allows for standard and fast cycling

✓ Compatible with all qPCR machines

Xpert Fast SYBR mastermix consists on the combination of a highly effi-cient enzyme, with a novel low inhibitory technology. The intercalating dye used in this mastermix causes little to no inhibition of the reaction thus allowing for extremely high sensitivity and specificity, as well as preventing the formation of unwanted primer-dimers and non-specific products.


GE20.0100 Xpert Fast SYBR (Uni) 1mL

GE20.5100 Xpert Fast SYBR (Uni) 5x 1mL

GE21.0100 Xpert Fast SYBR (Fluorescein)

1mL

GE21.5100 Xpert Fast SYBR (Fluorescein)

5x 1mL

Applications

Absolute quantification

Relative gene expression analysis

High throughput qPCR

Low-copy number target genes

Amplification of ACTG1 gene

Amplification of CANX gene

Amplification of GADPH gene

Amplification of PGK-1 gene

MX3000P®MX3005P®MX4000P®

CFX384™

Cromo4™ MiniOpticon™

Opticon™ Opticon™2

iCycler® iQ™5

MyiQ™Eco™

ABI 7000ABI 7300StepOne™

StepOne™Plus

ABI 7500ABI 7500 Fast

QuantStudio™ 12k FlexViiA7™

ABI 7700ABI 7900

ABI 7900HTABI 7900HT Fast

Rotor-Gene™ 3000Rotor-Gene™ 6000

Rotor-Gene™ Q

LightCycler® 480LightCycler® 96

LightCycler® Nano

Agilent BioRad Illumina Life Technologies Qiagen Roche

Xpert Fast SYBR (Uni)

Xpert Fast SYBR (Fluorescein)

Xpert Fast Probe (Uni)

Low ROX

Low ROX

No ROX

No ROX

No ROX

No ROX

No ROX

No ROX

High ROX

High ROX

DNA Amplification / qPCR


www.grisp.pt

43

Xpert Fast Probe ✓ High efficiency in multiplex reactions

✓ High efficiency in GC / AT rich templates

✓ Early Ct values - Rapid extension rate

✓ Extreme sensitivity – increased limit of detection

✓ Allows for standard and fast cycling

Xpert Fast Probe mastermix includes a highly efficient enzyme, combined with a novel buffer system. The performance of this master-mix allows for extremely high sensitivity and specificity, both for single-plex and multiplex applications, thus making this a robust option for confident probe qPCR results.


GE30.0100 Xpert Fast Probe (Uni) 1mL

GE30.5100 Xpert Fast Probe (Uni) 5x 1mL

Applications

Absolute quantification

Relative gene expression analysis

Low-copy number target genes

Multiplex or singleplex

Diagnostic real-time PCR

Amplification of ACVR1B gene

Amplification of ACVR2B gene

MX3000P®MX3005P®MX4000P®

CFX384™

Cromo4™ MiniOpticon™

Opticon™ Opticon™2

iCycler® iQ™5

MyiQ™Eco™

ABI 7000ABI 7300StepOne™

StepOne™Plus

ABI 7500ABI 7500 Fast

QuantStudio™ 12k FlexViiA7™

ABI 7700ABI 7900

ABI 7900HTABI 7900HT Fast

Rotor-Gene™ 3000Rotor-Gene™ 6000

Rotor-Gene™ Q

LightCycler® 480LightCycler® 96

LightCycler® Nano

Agilent BioRad Illumina Life Technologies Qiagen Roche

Low ROX

Low ROX

No ROX No ROX

No ROX No ROX

High ROX

High ROX

DNA Amplification / qPCR

44

GRS dNTP mix

GRS PCR Grade Water

GRS dNTP Set

Aqueous solution of dATP, dTTP, dCTP, and dGTP (10 mM each) at pH 7.0

Application(s)

For use in molecular biology applications, including PCR, long PCR, high fidelity PCR, real-time PCR, RT-PCR, cDNA synthesis, primer extension, and DNA labelling

Quality

✓ Functionally tested by PCR with Taq and Pfu polymerases

✓ Purity (>99%) confirmed by HPLC

✓ Free of endonuclease, exonuclease, RNase, and phosphatase activity

Application(s)

For use in molecular biology applications, including PCR, real-time PCR, RT-PCR.

Quality

✓ Free of endonuclease, exonuclease, RNase, and phosphatase activity.

✓ Free of nucleic acids.

✓ Not DEPC-treated

The set consists of 4x0,25ml, 100mM aqueous solutions of dATP, dTTP, dCTP, and dGTP, each supplied in a separate vial.

Application(s)

For use in molecular biology applications, including PCR, long PCR, high fidelity PCR, real-time PCR, RT-PCR, error-prone PCR, cDNA synthesis, primer extension, and DNA labelling

Quality

✓ Free of endonuclease, exonuclease, RNase, and phosphatase activity

✓ Functionally tested by PCR with Taq and Pfu polymerases dATP λmax=259nm; ε=15,2x103 M-1cm-1 at pH 7.0 dGTP λmax=253nm; ε=13,7x103 M-1cm-1 at pH7.0 dCTP λmax=271nm; ε=9,3x103 M-1cm-1 at pH 7.0 dTTP λmax=267nm; ε=9,6x103 M-1cm-1 at pH 7.0

✓ Purity (>99%) confirmed by HPLC

10mM each (min.99%)

100mM each (min.99%)

Ultrapure water


GP010.0001 GRS dNTP Mix(10mM each)

1mL

GP010.0501 GRS dNTP Mix(10mM each)

5x 1mL


GP011.0411 GRS dNTP Set(100mM each)

4x0,25mL


GW010.1001 GRS PCR Grade Water 10x1ml

GW010.1000 GRS PCR Grade Water 1L

DNA Amplification / dNTPs & Water


www.grisp.pt

45

Ultrapure water


GD02.0360 GRS PCR strips 0.2mL (attached flat caps)

360 units


GD03.0050 GRS 96w qPCR plates (half-skirted) 50 units


GD04.0100 GRS qPCR seals 100 units


GD01.5000 GRS 0.2mL PCR tubes (flat cap) 5000 units

GRS PCR PlasticsImprove your PCR with high quality PCR plastics. Performance of your PCR not only depends on the nature of the sample, the type of polymerase, and equipment, but is greatly affected by the quality of the plasticware as well. GRS PCR Plastics are certified free of DNase, RNase and human genomic DNA and manufactured of ultra pure polypropylene that inhibits protein (polymerase) binding to the surface. Very thin walls ascertain fast and homogenous heat transfer without compromising robust-ness. Moreover, GRS PCR Plastics demonstrate insignificant evaporation, which is especially important when performing small volume PCR reactions.

GRS 0,2ml PCR tubes (flat cap)

✓ Flat cap and frosted area for easy labeling

✓ Compatible with almost all thermocyclers

GRS 96w qPCR plates (half-skirted)

✓ 0,3ml reaction volume

✓ Dark printed alpha numeric code for easy identification

✓ Raised well rims

✓ compatible with GRS qPCR seals

GRS qPCR seals

✓ Easy removable clear seals, which leave no sticky residue once peeled off.

✓ suitable for all qPCR applications as well as for other fluorescence measurements.

✓ Compatible with PE, PP, and PS plates (135 x 80mm).

✓ Insignificant evaporation during cycling or storage

✓ Temperature range: -80°C to +120°C

GRS PCR strips 0,2ml (attached flat caps)

✓ Strips of 8 tubes with attached flat caps

✓ Flat caps and frosted area for easy labeling

✓ Compatible with almost all thermocyclers

DNA Amplification / PCR Plastics

Symon

PRODUCT CHANGE

Symon

PRODUCT CHANGE

Symon

PRODUCT CHANGE

Symon

PRODUCT CHANGE

RNA Research

cDNA Synthesis

GRS cDNA Synthesis Kit

GRS cDNA Synthesis Mastermix

PLUScript cDNA Synthesis Kit

GRS One-Step RT-PCR Kit

RNA Storage

RNA Stand-By Solution

Decontamination

RNase Xterminator Spray

48

48

49

50

51

51

04

48

GRS cDNA Synthesis KitKit supplied with separate components, including both oligo(dT) and random primers.

GRS cDNA Synthesis MastermixConvenient mastermix format, with separate oligo(dT) and random primers included.

The GRS cDNA Synsthesis Kit and GRS cDNA Synthesis Mastermix provide efficient and fast methods for the synthesis of first strand cDNA from purified poly(A)+ or total RNA templates.

Contain all required components needed to ensure high sensitivity, high yield, and full-length transcripts, and are suitable for first-strand cDNA synthesis, construction of cDNA libraries, probe generation for hybridization experiments, and primer extension. Moreover, first strand of cDNA synthesized with these kits can be used directly without purification as a template in PCR.

Contains

✓ GRS Reverse Transcriptase

✓ RNase inhibitor

✓ Oligo(dT)20 primer

✓ Random hexamer primer

Contains

✓ GRS Reverse Transcriptase MasterMix

✓ Oligo(dT)20 primer

✓ Random hexamer primer

✓ RNase-free water

(oligo dT)

(random hexamer)


GK24.0050 GRS cDNA Synthesis Kit 50 rxn

GK24.0100 GRS cDNA Synthesis Kit 100 rxn


GK34.0050 GRS cDNA Synthesis MasterMix 50 rxn

GK34.0100 GRS cDNA Synthesis MasterMix 100 rxn

Broad dynamic range

Thermostable and RNase H minus activity

Highly efficient synthesis of first strand cDNA

RNA Research / cDNA Synthesis

✓ dNTP mix

✓ 10x Reaction Buffer

✓ DTT



www.grisp.pt

49

Contains

✓ PLUScript RT/RNase inhibitor Mix

✓ cDNA 2x Reaction Buffer

✓ Anchored Oligo d(T)20 primer

✓ Random nonaprimer (N9)

✓ GRS gDNA Eraser


Conventional Oligo dT

Randomly binds to polyA tail

Anchored Oligo dT

Specifically binds adjacent to the first base

PLUScript cDNA Synthesis KitSample (total RNA):

Sample (mRNA): Operation Time:

50ng - 5ug

5-500ng

35 minutes

PLUScript Reverse Transcriptase is a genetically modified M-MLV reverse transcriptase with deficient RNase H activity, improved synthesis efficiency, and enhanced thermostability and thus suitable for GC-rich and complex secondary structure RNA templates, as first-strand cDNA synthesis can be performed at higher temperatures.

PLUScript RT is provided as a convenient enzyme mix containing RNase inhibitor and supplied with a complete reaction mix including DTT and dNTPs, allowing for less pipetting steps, minimizing contamination and reducing hands-on time. Furthermore, besides random primers (N9), the kit contains anchored oligo d(T)20 primers, which bind specifically to the 5´-beginning of the poly(A) tail resulting in an increased full-length cDNA yield as in comparison with traditional oligo(dT) primers.

By combining PLUScript cDNA Synthesis with GRS gDNA Eraser (inclu-ded), first-strand cDNA synthesis can be accomplished with simulta-neous genomic DNA removal in a single tube reaction. This eliminates the need of DNase I treatment of the RNA sample prior to cDNA synthe-sis, reducing both total and hands-on time. Moreover, as DNase I needs to be heat-inactivated, possible degradation of RNA is minimized.

PLUScript Reverse Transcriptase and GRS gDNA eraser are the perfect combination for first strand cDNA synthesis for demanding samples and is recommended for expression analysis of multiple copy and low copy genes, cDNA library construction, 3´- and 5´-RACE, and difficult RNA templates.

Very high efficiency

cDNA up to 15kb

Simultaneous cDNA Synthesis and gDNA removal

Ideal for GC-rich and Complex Secondary Struture RNA

Fast (35 minutes)


GK44.0050 PLUScript cDNA Synthesis Kit 50rxn


50

Contains

✓ GRS RT-PCR Enzyme Mix

✓ 2x GRS One-step RT-PCR Mastermix


GRS One-Step RT-PCR KitSample:Application:

1pg - 1ug RNAMultiple copy and low copy gene detection

Using gene-specific primers, the GRS One-Step RT-PCR Kit allows for first-strand cDNA synthesis and subsequent PCR in a single-tube reac-tion procedure, decreasing contamination risk and reducing hands-on time considerably.

The kit consist of an enzyme mix comprising a genetically modified M-MLV reverse transcriptase with deficient RNase H activity and impro-ved synthesis efficiency combined with RNase inhibitor, and a conve-nient mastermix containing all other required components, including a high-fidelity DNA polymerase blend, dNTPs and tracking dye allowing for direct loading onto agarose gels for subsequent DNA electrophore-sis. PCR products contain 3´ A-overhang and are thus suitable for TA-clo-ning. The kit allows for the amplification of fragments up to 8kb.

Β-actin amplification - Lanes 1-9: 0 pg, 0.1 pg, 1 pg, 10 pg, 100 pg, 1000 pg, 10 ng, 100ng, 1000 ng

GRS One-Step RT-PCR Kit

cDNA Synthesis & PCR

in one tube

+ Gene Specific Primers + RNA template


GK54.0100 GRS One-Step RT-PCR Kit 100rxn

Easy Reaction set-up

Little hands-on time

Single-tube reaction - less contaminations



www.grisp.pt

51


GB33.0100 RNA Stand-by Solution 100mL


GB43.500S RNase Xterminator Spray

500mL

RNA Stand-by Solution

RNase Xterminator Spray

Application:

Protecting RNA, especially in RNase-rich tissues Archiving tissues Collecting samples at various times without need of processing right away Shipping samples

RNA Stand-by Solution is an aqueous solution that inactivates RNases and preserves cellular RNA of intact fresh tissues or cells.

With RNA stand-by, you no longer need to rush in order to process your tissue samples immediately or freeze them in liquid nitrogen. Simply immerse your tissue sample in RNA Stand-by Solution and store as convenient, without putting the integrity of the RNA at risk. RNA Stand-by can be added to pieces of tissue, cell pellets and even culture medium. Using RNA Stand-by Solution does not jeopardize quality nor quantity of the RNA to be isolated subsequently., whether the sample is stored frozen or not.

RNA is stable for up to 1 day when stored at 37°C, up to 1 week at room temperature, and up to 1 month in a normal refrigerator. For long term, samples should be stored frozen at -20°C.

Perfect for tissue collection outside the lab

Flexible tissue storage

Compatible with most RNA purification methods

RNase Xterminator Spray is a ready-to-use solution, supplied in an easy-to-use Spray Bottle, for eliminating RNase and DNA contamination from laboratory surfaces. It can be used for the decontamination of DNA, RNase and other enzymes from labware, equipment, and bench tops. Simply spray onto the contaminated area and wipe away using RNase-free water.

RNA Research / RNA Storage & Decontamination

Cloning

Cloning Kits

GRS TOP-TA positive Cloning Kit

GRS TOP-Blunt positive Cloning Kit

Miscelaneous

T4 DNA Ligase

IPTG

X-Gal

Antibiotics

Ampicillin

Carbenecillin

Chloramphenicol

Gentamycin

Kanamycin

Tetracycline

54

55

56

56

56

57

57

57

57

57

57

Restriction Enzymes

Xpress BamHI

Xpress EagI

Xpress EcoRI

Xpress EcoRV

Xpress HindIII

Xpress NcoI

Xpress NdeI

Xpress NheI

Xpress NotI

Xpress PstI

Xpress ScaI

Xpress SmaI

Xpress SphI

Xpress XbaI

Xpress XhoI

Xpress XmaI

58

58

58

58

58

58

58

58

58

58

58

58

58

58

58

58

05

54

pGRS series map

Cloning site pGRS TOP-TA

Cloning site pGRS TOP-Blunt

Spe I

Pst I

M13 Reverse Primer

Pme I Not I Apo I

T3 Promoter

T7 Promoter M13 Forward Primer

PCR product w/ 3´A-overhang

Spe I

Pst I

M13 Reverse Primer

PCR productblunt

Pme I Not I Apo I

T3 Promoter

T7 Promoter M13 Forward Primer

Cloning / Cloning Kits


www.grisp.pt

55

GRS TOP-TA Positive Cloning KitThe GRS TOP-TA positive cloning kit is designed for the ligation of PCR fragments containing 3´-A overhangs (obtained using “normal” Taq) into the plasmid disrupting the expression of a lethal gene. Upon transfor-mation, only E.coli harbouring recombinant vector survive and there is no need for blue/white screening. Ligation can be done in as little as 5 minutes with high efficiency. The linearized pGRS TOP-TA vector contains ampicillin and kanamycin resistance genes for selection. The PCR amplicon is inserted between M13 primers (without the requirement of DNA Ligase) which facilitate DNA sequencing. Flanking Lac, T3, and T7 promoters ensure the possibility for correct in vitro transcription of the insert independent of its orientation.

GRS TOP-Blunt Positive Cloning KitThe GRS TOP-Blunt positive cloning kit is designed for the ligation of blunt-ended PCR fragments (obtained using high-fidelity polymeras-es such as our Xpert HighFidelity DNA polymerase) into the plasmid disrupting the expression of a lethal gene. Upon transformation, only E.coli harbouring recombinant vector survive and there is no need for blue/white screening. Ligation can be done in as little as 5 minutes with high efficiency and without the requirement of adding DNA Ligase. The linearized pGRS TOP-Blunt vector contains ampicillin and kanamycin resistance genes for selection. The PCR amplicon is inserted between M13 primers, which facilitate DNA sequencing. Flanking Lac, T3, and T7 promoters ensure the possibility for correct in vitro transcription of the insert independent of its orientation.

Features

✓ Fast Ligation, no need for DNA Ligase

✓ Positive selection, no need for blue-white screening

✓ High Efficiency, virtually 100% positive clones

✓ Flanking M13 primers for Sequencing

✓ Flanking promoters for correct in vitro transcription

Components

✓ Linearized vector (10ng/μL)

✓ M13 primers

✓ Control insert

✓ Control primer

GRS positive cloning vectors are designed for the ligation of PCR fragments into the plasmid disrupting the expression of a lethal gene. Upon transformation, E.coli harbouring non-recombinant vector die whilst cells harbouring recombinant vector containing the desired insert survive. Virtually all clones are positive and, therefore, there is no need for blue/white screen-ing. GRS positive cloning kits contain all components necessary for the rapid ligation of PCR fragments for both sequencing and in vitro transcription, without the requirement of DNA Ligase. The GRS TOP-TA Cloning kit is suitable for PCR ampli-cons obtained using “normal” Taq polymerase, which creates 3´-A overhangs , whereas the GRS TOP-Blunt Cloning kit is designed for PCR products obtained with high-fidelity polymerases, which result in blunt-ended PCR fragments.


GC01.0020 GRS TOP-TA Positive Cloning Kit 20 rxn


GC02.0020 GRS TOP-Blunt Positive Cloning Kit 20 rxn

Cloning / Cloning Kits

56

T4 DNA Ligase (5U/μL)T4 DNA Ligase catalyzes the formation of phosphodiester bonds between adjacent 5´- phosphate and 3´-hydroxyl termini in double-stranded DNA and/or double-stranded RNA. T4 DNA Ligase is capable of joining blunt and complementary cohesive ends, making it the Ligase of choice for restriction-ligation cloning of DNA fragments into plasmids. Moreover, this enzyme can be used for nick-repair as it closes nicks in double-stranded DNA or DNA/RNA hybrids. GRiSP´s T4 DNA Ligase has a concentration of 5 Weiss units per microliter and is supplied with a 5x reaction buffer.

IPTG (max 5ppm dioxane)Isopropyl ß-D-thiogalactopyranosideIPTG (Isopropyl ß-D-thiogalactopyranoside) is a lactose analog that mimics allolactose, which triggers transcription of the lac operon by inactivation of the lac repressor. As such, IPTG can be used to induce protein expression of heterologous proteins that are encoded by genes under the control of the lac operator. Unlike allolactose, IPTG is not hydrolyzed by ß-galactosidase, and its concentration remains constant during protein expression experiments. IPTG is effective at concentra-tions ranging from 0.1-1.0 mM. In combination with X-gal, IPTG can be used for blue/white screening of the lac phenotype of E.coli in cloning experiments and discriminate between bacterial colonies transformed with recombinant plasmid containing the DNA fragment of interest (white) versus non-recombinant plasmid (blue). GRiSP´s IPTG is at least 99% pure and has a very low content of dioxane, a compound that has a negative impact on normal cell function.

X-Gal5-Bromo-4-chloro-3-indolyl-ß-D-galactopyranosideX-Gal (5-Bromo-4-chloro-3-indolyl-ß-D-galactopyranoside) is a synthetic analog of lactose that mimics lactose as an inert chromogenic substrate for ß-galactosidase. In combination with, IPTG (needed for the induc-tion of the expression of ß-galactosidase), X-Gal can be used for blue/white screening of the lac phenotype of E.coli in cloning experiments and discriminate between bacterial colonies transformed with recom-binant plasmid containing the DNA fragment of interest (white) versus non-recombinant plasmid (blue), as ß-galactosidase hydrolyzes X-gal into galactose and an indigo-coloured precipitate. GRiSP´s X-gal is at least 99% pure as determined by HPLC.


GC03.1000 T4 DNA Ligase (5U/μL) 1000U


GAB01.0005 IPTG (max 5ppm dioxane) 5g


GAB02.0005 X-Gal 5g

Cloning / Miscelaneous


www.grisp.pt

57

Antibiotics are used in a wide range of molecular biology applications including cloning experiments. Upon transformation, incubation using a medium supplemented with a specific antibiotic allows for the selection of bacterial cells harbouring vectors containing genes that confer resistance to that specific antibiotic, as untransformed cells are sensitive and die. GRiSP offers a set of the most commonly used antibiotics for molecular cloning. Please check both your plasmid map and E.coli genotype to select the appropriate antibiotic(s).

Ampicillin

GentamycinTetracycline

Carbenicillin Kanamycin

ChloramphenicolChloramphenicol is a bacteriostatic agent that acts by binding to the 50S subunit inhibiting peptidyl transferase required for protein synthesis.A stock solution of 10mg/ml should be prepared using methanol and stored at 4°C. Recommended final concentration is 10-25µg/ml. Purity (on dried basis): min 98%


GAB03.0005 Ampicillin (sodium salt) 5g


GAB04.0005 Kanamycin (sulphate) 5g


GAB07.0005 Tetracycline (hydrochloride) 5g


GAB08.0005 Gentamycin (sulphate) 5g


GAB06.0005 Carbenicillin (disodium salt) 5g


GAB05.0005 Chloramphenicol 5g

Sodium salt

SulphateHydrochloride

Disodium salt Sulphate

Ampicillin is a bacteriocidal agent that acts by inhibition of transpeptidase, which is required for cell wall synthesis. It is recommended to prepare a stock solution of 4mg/ml in sterile, ultrapure water, which should be stored at 4°C. Recommended final concentration is 50-100µg/ml. Purity (on dried basis): min 90%.

Gentamycin is an aminoglycoside antibiotic complex com-prised of 3 major components (C1, C1a, and C2). It inhibits protein synthesis by binding to the 30S subunit of the ribo-some. A stock solution of 10mg/ml in sterile ultrapure water can be stored at 4°C. Recommended final concentration is 10-15µg/ml. Activity (on dried basis): min 590 ug/mg.

Tetracyclin is a bacteriostatic agent that prevents aminoacyl tRNA from biding to the ribosome A site. It is recommend-ed to prepare a stock solution of 12mg/ml in 70% ethanol, which should be stored at -20°C. Recommended final concentration is 12µg/ml.Tetracyclin is light sensitive and product and medium con-taining tetracycline should be stored in the dark.

Carbenicillin, like ampicillin, is an antibiotic of the ß-lactam class and acts in an identical way as ampicillin and has the advantage that it is more stable. Prepared plates and broth can be stored for longer period than with ampicil-lin, and there is less risk of formation of satellite-colonies. Purity (on dried basis): min 90%.

Kanamycin is a bacteriocidal agent that binds to the 30S ribosomal subunit inhibiting protein synsthesis. It is recom-mended to prepare a stock solution of 10mg/ml in sterile, ultrapure water, which should be stored at 4°C. Recommended final concentration is 30-100µg/ml. Potency: min 750Ug/mg

Composition Gentamycin : C1a: 10 - 35%C2: 25 - 55%C1: 25 - 50%

Cloning / Antibiotics

58

Xpress Restriction EnzymesGRiSP offers a set of fast restriction enzymes for cloning experiments that demonstrate high efficiency in an universal buffer. Complete diges-tion can be achieved in as little as 5 minutes whereas prolonged diges-tion does not result in any detectable star activity. All our restriction endonucleases can be heat-inactivated, therefore one can proceed immediately to the ligation step.

Reference Description Amount Recognition Site Heat Inactivation Concentration

GR01.5000 Xpress BamHI 5000 units 5'... G↓GATCC …3' 20 min at 65ºC 20,000 U / mL *available soon

GR02.0250 Xpress EagI 250 units 5'... C↓GGCCG …3' 20 min at 65ºC 10,000 U / mL

GR03.5000 Xpress EcoRI 5000 units 5'... G↓AATTC …3' 20 min at 65ºC 20,000 U / mL

GR04.2000 Xpress EcoRV 2000 units 5'... GAT↓ATC …3' 20 min at 65ºC 20,000 U / mL

GR05.5000 Xpress HindIII 5000 units 5'... A↓AGCTT …3' 20 min at 65ºC 20,000 U / mL

GR06.0500 Xpress NcoI 500 units 5'... C↓CATGG …3' 20 min at 80ºC 10,000 U / mL

GR07.2000 Xpress NdeI 2000 units 5'... CA↓TATG …3' 20 min at 65ºC 20,000 U / mL

GR08.0500 Xpress NheI 500 units 5'... G↓CTAGC …3' 20 min at 80ºC 10,000 U / mL

GR09.0250 Xpress NotI 250 units 5'... GC↓GGCCGC …3' 20 min at 65ºC 10,000 U / mL

GR10.5000 Xpress PstI 5000 units 5'... CTGCA↓G …3' 20 min at 80ºC 20,000 U / mL

GR11.1000 Xpress ScaI 1000 units 5'... AGT↓ACT …3' 20 min at 80ºC 20,000 U / mL *available soon

GR12.1000 Xpress SmaI 1000 units 5'... CCC↓GGG …3' 20 min at 65ºC 20,000 U / mL

GR13.0500 Xpress SphI 500 units 5'... GCATG↓C …3' 20 min at 65ºC 20,000 U / mL

GR14.1500 Xpress XbaI 1500 units 5'... T↓CTAGA …3' 20 min at 65ºC 20,000 U / mL

GR15.2500 Xpress XhoI 2500 units 5'... C↓TCGAG …3' 20 min at 65ºC 20,000 U / mL

GR16.0250 Xpress XmaI 250 units 5'... C↓CCGGG …3' 20 min at 65ºC 10,000 U / mL

Cloning / Restriction Enzymes


www.grisp.pt

59

Xpress Starter Pack available, including: 5000U Xpress PstI, 5000U Xpress EcoRI, 2000U Xpress EcoRV, 250U Xpress NotI, 1500U Xpress XbaI)

Reference: GR00.SP1

Reference Description Amount Recognition Site Heat Inactivation Concentration

GR01.5000 Xpress BamHI 5000 units 5'... G↓GATCC …3' 20 min at 65ºC 20,000 U / mL *available soon

GR02.0250 Xpress EagI 250 units 5'... C↓GGCCG …3' 20 min at 65ºC 10,000 U / mL

GR03.5000 Xpress EcoRI 5000 units 5'... G↓AATTC …3' 20 min at 65ºC 20,000 U / mL

GR04.2000 Xpress EcoRV 2000 units 5'... GAT↓ATC …3' 20 min at 65ºC 20,000 U / mL

GR05.5000 Xpress HindIII 5000 units 5'... A↓AGCTT …3' 20 min at 65ºC 20,000 U / mL

GR06.0500 Xpress NcoI 500 units 5'... C↓CATGG …3' 20 min at 80ºC 10,000 U / mL

GR07.2000 Xpress NdeI 2000 units 5'... CA↓TATG …3' 20 min at 65ºC 20,000 U / mL

GR08.0500 Xpress NheI 500 units 5'... G↓CTAGC …3' 20 min at 80ºC 10,000 U / mL

GR09.0250 Xpress NotI 250 units 5'... GC↓GGCCGC …3' 20 min at 65ºC 10,000 U / mL

GR10.5000 Xpress PstI 5000 units 5'... CTGCA↓G …3' 20 min at 80ºC 20,000 U / mL

GR11.1000 Xpress ScaI 1000 units 5'... AGT↓ACT …3' 20 min at 80ºC 20,000 U / mL *available soon

GR12.1000 Xpress SmaI 1000 units 5'... CCC↓GGG …3' 20 min at 65ºC 20,000 U / mL

GR13.0500 Xpress SphI 500 units 5'... GCATG↓C …3' 20 min at 65ºC 20,000 U / mL

GR14.1500 Xpress XbaI 1500 units 5'... T↓CTAGA …3' 20 min at 65ºC 20,000 U / mL

GR15.2500 Xpress XhoI 2500 units 5'... C↓TCGAG …3' 20 min at 65ºC 20,000 U / mL

GR16.0250 Xpress XmaI 250 units 5'... C↓CCGGG …3' 20 min at 65ºC 10,000 U / mL

Features

✓ Fast 5 minute digestion

✓ One buffer for all options

✓ No star activity

✓ Highly efficient

✓ Supplied with (separate) Loading Buffer

Culture Media

Auto Induction Media

LB Broth (AIM)

2x YT Broth (AIM)

Terrific Broth (AIM)

Super Broth (AIM)

Standard Media

LB Agar (Lennox)

LB Broth (Lennox)

Luria Agar (Miller’s LB Agar)

Luria Broth (Miller’s LB Broth)

Luria Agar (Miller’s Modification)

Luria Broth (Miller’s Modification)

Terrific Broth

Modified Terrific Broth

2x YT Medium

62

62

62

62

63

63

63

63

63

63

63

63

63

2xYT Agar

SOB medium

SOC Medium

YPD Agar

YPD Broth

YNB w/o aa, w/o ammonium sulphate

YNB w/o aa, with ammonium sulphate

Media Components

Peptone

Bacteriological Peptone

Tryptone

Yeast Extract

Bacteriological Agar

Dextrose

Sucrose

0663

63

63

63

63

63

63

62

62

62

62

62

62

62

62

Auto Induction MediaDehydrated powders, supplemented with glucose and alpha lactose for the auto induction of protein expression under the control of IPTG-inducible promoters in Escherichia coli. Media contain per L 0,15g MgSO4, 3,3g (NH4)2SO4, 7,1g Na2HPO4, and 6,8g KH2PO4.

Variable components, Tryptone and Yeast Extract, between the different options are shown in the table below.

✓ No cell density monitoring needed

✓ Automatic induction of protein expression


GCM17.0500 LB Broth (AIM) 500gTryptone: 10g/L | Yeast Extract: 5g/L

GCM18.0500 2x YT Broth (AIM) 500gTryptone: 16g/L | Yeast Extract: 10g/L

GCM19.0500 Terrific Broth (AIM) 500gTryptone: 12g/L | Yeast Extract: 24g/L

GCM20.0500 Super Broth (AIM)

Tryptone: 35g/L | Yeast Extract: 20g/L

500g

Media ComponentsComponents for the preparation of commonly used culture media in molecular biology experiments.


GCM21.0500 Peptone 500g

GCM22.0500 Bacteriological Peptone 500g

GCM23.0500 Tryptone 500g

GCM24.0500 Yeast Extract 500g

GCM25.0500 Bacteriological Agar 500g

GCM26.0500 Dextrose 500g

GCM27.0500 Sucrose 500g

Cultura Media / AIM Media & Media Components


www.grisp.pt

63

Standard MediaDehydrated powder for the preparation of broth or agar plates, for the groth of bacteria or yeast in molecular biology experiments.


GCM01.0500 LB Agar (Lennox) 500gTryptone: 10g/L | Yeast Extract: 5g/L | NaCl: 5g/L | Bacteriological Agar: 15g/L

GCM02.0500 LB Broth (Lennox) 500gTryptone: 10g/L | Yeast Extract: 5g/L | NaCl: 5g/L

GCM03.0500 Luria Agar (Miller's LB Agar) 500gTryptone: 10g/L | Yeast Extract: 5g/L | NaCl: 10g/L | Bacteriological Agar: 15g/L

GCM04.0500 Luria Broth (Miller's LB Broth) 500gTryptone: 10g/L | Yeast Extract: 5g/L | NaCl: 10g/L

GCM05.0500 Luria Agar (Miller's Modification) 500gTryptone: 10g/L | Yeast Extract: 5g/L | NaCl: 0,5g/L | Bacteriological Agar: 15g/L

GCM06.0500 Luria Broth (Miller's Modification) 500gTryptone: 10g/L | Yeast Extract: 5g/L | NaCl: 0,5g/L

GCM07.0500 Terrific Broth 500gTryptone: 12g/L | Yeast Extract:24g/L | K2HPO4: 12,54g/L | KH2PO4: 2,31g/L

GCM08.0500 Modified Terrific Broth 500gTryptone: 12g/L | Yeast Extract:24g/L | K2HPO4: 9,40g/L | KH2PO4: 2,20g/L

GCM09.0500 2x YT Medium 500gTryptone: 16g/L | Yeast Extract: 10g/L | NaCl: 5g/L

GCM10.0500 2x YT Agar 500gTryptone: 16g/L | Yeast Extract: 10g/L | NaCl: 5g/L | Bacteriological Agar: 15g/L

GCM11.0500 SOB medium 500gTryptone: 20g/L | Yeast Extract: 5g/L | NaCl: 0,5g/L | MgSO4: 0,96g/L | KCl: 0,186g/L

GCM12.0500 SOC Medium 500gTryptone: 20g/L | Yeast Extract: 5g/L | NaCl: 0,5g/L | MgSO4: 0,96g/L | Glucose: 3,60g/L | KCl: 0,186g/L

GCM13.0500 YPD Broth 500gPeptone: 20g/L | Yeast Extract: 10g/L | Dextrose: 20g/L

GCM14.0500 YPD Agar 500gPeptone: 20g/L | Yeast Extract: 10g/L | Dextrose: 20g/L | Bacteriological Agar: 15g/L

GCM15.0500 YNB without amino acids and without ammonium sulfate 500gYNB: 1,7g/L

GCM16.0500 YNB without amino acids and with ammonium sulfate 500gYNB: 1,7g/L | Ammonium sulfate: 5g/L

Cultura Media / Standard Media

Protein Research

Acrylamide / Bisacrylamide Solutions

Acrylamide / Bisacrylamide – 30%

Acrylamide / Bisacrylamide – 40%

Solutions

TG Buffer

TGS Buffer

SDS Solution

Protein Markers

GRS Unstained Protein Marker

GRS Protein Marker – Blue

GRS Protein Marker - MultiColour

GRS Protein Marker – MultiColour Plus

66

66

67

67

67

68

68

69

69

Staining & Stripping

Ponceau S Solution

GRS Stripping Solution

Quantification

GRS Protein Quantification Kit (Bradford)

GRS Protein Quantification Kit (BCA)

0770

70

71

71

6666


GB16.3019 Acrylamide/Bisacrylamide Solution 30%(19:1) 500mL


GB16.3037 Acrylamide/Bisacrylamide Solution 30%(37.5:1) 500mL

Acrylamide-Bisacrylamide SolutionsConcentrated mixtures of acrylamide and bisacrylamide for the preparation of polyacrylamide gels.Features

Ready-to-Use Acrylamide-Bisacrylamide solutions are aqueous solutions of ultra-pure acrylamide and bisacrylamide prepared with ultrapure water, 0.2µm filtered, and dispensed in an amber plastic bottle.

Pore Size

The range of protein sizes to be separated is determined by the pore size of a polyacrylamide gel. Control of the pore size is accomplished by regulating the total (T) amount of acrylamide as well as the percentage of cross-linker (C). The relationship between T and pore size is nearly linear. Gels with a higher percentage T have smaller pores, and are used to separate smaller proteins. The relationship between C and pore size is more complex. Pore size decreases with increasing C with a minimum occurring at about 5%. Increasing C further results in increas-ing pore size due to of non-homogeneous bundling of polymer strands.




GB16.4037 Acrylamide/Bisacrylamide Solution 40%(37.5:1) 500mL

Protein Reseach / Acrylamide - Bisacrylamide Solutions


www.grisp.pt

67www.grisp.ptalways here for you

www.grisp.pt

67

TGS Buffer (10x)Tris-Glycine-SDS Running Buffer, 10x concentratedTris-glycine-SDS (TGS) running buffer is the most common buffer used for sodium dodecyl sulfate – polyacrylamide gel electrophoresis of proteins (SDS-PAGE).

Features

TGS Buffer (10x) is a 0.2µm filtered aqueous solution of 0,25M Tris, 1,92M glycine, and 1% SDS prepared with ultrapure water. It is supplied in a 1L plastic bottle or in a 5L handy Bag-in-Box equipped with a two-position spigot that offers push button dispensing.

TG Buffer (10x)Tris-Glycine Running Buffer, 10x concentratedTris-glycine (TG) running buffer is a common buffer used for non-dena-turing (native) gel electrophoresis of proteins and for protein transfer in Western blotting.

Features

TG Buffer (10x) is a 0.2µm filtered aqueous solution of 0,25M Tris, and 1,92M glycine, prepared with ultrapure water. It is supplied in a 1L plastic bottle or in a 5L handy Bag-in-Box equipped with a two-position spigot that offers push button dispensing.

SDS solutionSDS solutions, 10% or 20%10% or 20% (w/v) solutions of sodium dodecyl sulfate prepared with ultrapure water, 0,2 μm filtered, and dispensed into 1L plastic bottles.


GB15.0110 TGS buffer 10X 1L

GB15.0510 TGS buffer 10X 5L


GB13.0110 TG buffer 10X 1L

GB13.0510 TG buffer 10X 5L


GB14.0110 SDS Solution 10% 1L

GB14.0120 SDS Solution 20% 1L

Protein Reseach / Solutions

6868

GRS Unstained Protein MarkerRecommended Loading:3-5 μL/lane (depending on the thickness of the gel) for staining with Coomassie Brilliant Blue (100-150 lanes)The GRS Unstained Protein Marker is a ready-to-use unstained protein standard suitable for size determination of proteins. It is composed of 8 unstained proteins and 1 pre-stained protein (blue). The unstained proteins range from 20kDa to 120kDa, whereas the pre-stained protein co-migrates with proteins of approximately 12kDa (depending on the SDS-PAGE conditions). The presence of the pre-stained band allows for monitoring electrophoresis and Western Blotting onto membrane (compatible with PVDF, nylon and nitrocellulose). For easy identifica-tion of each band, the 50kDa band has double intensity to serve as an internal reference.

GRS Protein Marker BlueRecommended Loading: 3-5 μL/lane (depending on the thickness of the gel) for clear visualization during elec-trophoresis (~125 lanes) 1,5-2,5 μL/lane for Western Blotting (~250 lanes)The GRS Protein Marker Blue is a ready-to-use blue protein standard suitable for monitoring protein separation during SDS-PAGE, size deter-mination of proteins, and verification of transfer efficiency of Western Blotting onto membrane (compatible with PVDF, nylon and nitrocellu-lose). This marker is composed of 11 pre-stained proteins ranging from 10kDa to 180kDa. For easy identification of each band, it includes two reference bands with increased intensity that co-migrate with proteins of 25kDa and 72kDa, respectively, when separated by 10% SDS-PAGE using TGS Buffer.

kDa

12010080605040

30

20

12 (pre-stained)

Tris-Glycine12%


GLP10.0500 GRS Unstained Protein Marker 500uL

GLP10.3500 GRS Unstained Protein Marker 3x 500uL


GLP02.0500 GRS Protein Marker Blue (100 lanes) 500uL

GLP02.3500 GRS Protein Marker Blue (300 lanes) 3x 500uL

Tris-Glycine10%

kDa

~180~140

~100

~72

~60

~45

~35

~25~20~15~10

Protein Reseach / Protein Markers


www.grisp.pt


www.grisp.pt

69

GRS Protein Marker MultiColourRecommended Loading:3-5 μL/lane (depending on the thickness of the gel) for clear visualization during elec-trophoresis (~125 lanes) 1,5-2,5 μL/lane for Western Blotting (~250 lanes)The GRS Protein Marker MultiColour is a ready-to-use three-colour protein standard suitable for monitoring protein separation during SDS-PAGE, size determination of proteins, and verification of transfer efficiency of Western Blotting onto membrane (compatible with PVDF, nylon and nitrocellulose). This marker is composed of 12 pre-stained proteins ranging from 10kDa to 245kDa. For easy identification of each band, it includes two reference bands of different colours that co-mi-grate with proteins of 25kDa (green) and 75kDa (red), respectively, when separated by 4-20% gradient SDS-PAGE using TGS Buffer.

GRS Protein Marker MultiColour PLUSRecommended Loading: 3-5 μL/lane (depending on the thickness of the gel) for clear visualization during elec-trophoresis (~125 lanes) 1,5-2,5 μL/lane for Western Blotting (~250 lanes)The GRS Protein Marker MultiColour PLUS is a ready-to-use three-co-lour protein standard suitable for monitoring protein separation during SDS-PAGE, size determination of proteins, and verification of transfer efficiency of Western Blotting onto membrane (compatible with PVDF, nylon and nitrocellulose). This marker is composed of 10 pre-stained proteins ranging from 6,5kDa to 270kDa. For easy identification of each band, it includes three reference bands of different colours that co-mi-grate with proteins of 30kDa (red), 52kDa (green) and 270kDa (red), respectively, when separated by 4-20% gradient SDS-PAGE using TGS Buffer.


GLP01.0500 GRS Protein Marker MultiColour (100 lanes) 500uL

GLP01.3500 GRS Protein Marker MultiColour (300 lanes) 3x 500uL


GLP03.0500 GRS Protein Marker MultiColour PLUS (100 lanes) 500uL

GLP03.3500 GRS Protein Marker MultiColour PLUS (300 lanes) 3x 500uL

kDa

~245~180~135~100~75~63

~48

~35

~25~20~17

~11

Tris-Glycine4 - 20%

kDa

~270~175~130

~95~66~52

~37

~30

~16

~6.5

Tris-Glycine4 - 20%

Protein Reseach / Protein Markers

7070

Ponceau S SolutionPonceau S Solution allows for the rapid and reversible detection of proteins on nitrocellulose and PVDF membranes for the verification of the transfer efficiency of Western Blotting before proceeding with incubation with primary antibody. Transfer artifacts, such as uneven transfer or bubbles, can be quickly detected, preventing continuation with imperfect membranes, saving you time and expen-sive antibodies. Ponceau S does not damage the antigen or has any deleterious effects on the sequence of blotted proteins and polypeptides and is therefore the method of choice for location for blot-sequencing. Staining can be carried out in less than 5 minutes and destaining also takes only a few minutes of incu-bation in water. After destaining, one can proceed immediately with subsequent steps of the Western Blotting protocol

GRS Stripping SolutionGRS Stripping Solution is designed for removing antibodies from developed membranes after Western Blotting, allowing for multiple detection with other sets of antibodies (reprobing).

The GRS Stripping Solution weakens protein-protein interactions and therefore removes primary and secondary antibodies from membranes, whilst transferred proteins remain on the membrane and the membrane itself undamaged. The solution does not contain DTT or β-mercaptoeth-anol, thus leaving disulfide bridges intact. The GRS Stripping Solution cannot be used with colourimetric substrates that precipitate (e.g. DAB or BCIP/NBT) but is intended for chemiluminescence or fluorescence detection.


GB21.0500 Ponceau S Solution 500mL


GB20.0500 GRS Stripping Solution 500mL

✓ Save time

✓ Save antigen

Protein Reseach / Staining & Stripping


www.grisp.pt


www.grisp.pt

71

GRS Protein Quantification Kit (Bradford)The Bradford protein assay is one of the most commonly used methods to determine total protein concentration. This colorimet-ric assay, based on binding of Coomassie Brilliant Blue G-250 to proteins, which leads to an absorbance shift of the dye, is very easy to set up, fast, and compatible with protein solutions contain-ing reducing agents. Change of colour can be observed within 5 to 10 minutes, with best results obtained within 20 minutes. Easy to use: One solution, no further preparation required

Fast: Within minutes, with optimum after about 20 min

Broad Range: Standard assay: 200-1500 µg/ml, micro assay 1-10 µg/ml

Components

✓ 100 ml of Coomassie Brilliant Blue G-250 Solution

✓ 4x1ml of BSA standard solution (0,22mg/ml)

GRS Protein Quantification Kit (BCAThe BCA protein assay is one of the most commonly used methods to determine total protein concentration. This colorimetric assay com-bines the biuret method with the chelating properties of the chro-mogenic reagent bicinchoninic acid and is highly sensitive as well as compatible with detergent solubilized protein solutions.

Compared to Coomassie dye-binding methods (Bradford), the BCA protein assay is less affected on the amino acid sequence/composition, providing greater consistency between different proteins.

The GRS Protein Quantification Kit (BCA) can be used for the quantifica-tion of total protein in a wide variety of samples, containing up to 5% of either one of the detergents SDS, Triton X-100, Tween®20, Tween®60, Tween®80, NP 40, or CHAPS.

Sensitive:detection limit of approximately 5µg/ml

Accurate:less protein-to-protein variation than Bradford

High Linearity:BSA: 20µg/ml to 2mg/ml

Components

✓ 100 ml of BCA Solution A

✓ 3ml of BCA solution B

✓ 2x 1ml of BSA standard solution (2 mg/ml)


GK51.0100 GRS Protein Quantification Kit (Bradford) 1 kit


GK50.0100 GRS Protein Quantification Kit (BCA) 1 kit

Protein Reseach / Quantification

Cell Biology

Cell Detachment

Accutase®

Trypsin / EDTA Solution

Supplements

L-Glutamine

Stable L-Glutamine

Penicillin-Streptomycin Solution

74

74

75

75

75

08

74

Accutase®Cell Detachment SolutionAccutase® is a ready-to-use non-mammalian, non-bacterial replacement for all applications of trypsin.

Accutase is a natural enzyme mixture with proteolytic and collagen- olytic enzyme activity. This means it mimics the action of trypsin and collagenase at the same time. However, because it is more efficient than mammalian trypsin & collagenase, it is formulated at a much lower concentration making it less toxic and more gentle.

✓ Can be used whenever gentle and efficient detachment of any adher-ent cell line is needed. Accutase is a direct replacement for trypsin.

✓ Works extremely well on embryonic and neuronal stem cells; mono layers of stem cells can be grown after passaging with Accutase®.

✓ Preserves most epitopes for subsequent flow cytometry analysis.

✓ Does not need to be neutralized when passaging adherent cells. The addition of more media after the cells are split dilutes Accutase® so it is no longer able to detach cells.

✓ Does not need to be aliquoted. A bottle is stable in the refrigerator for 2 months

Accutase performs exceptionally well in detaching cells for: hESC cultur- ing, analysis of cell surface markers, virus growth assay, quiescence assays by serum starvation, transformation assays by oncogene transfection, neural crest cell migration assays, cell proliferation, apopto- sis, cell haptotaxsis, tumor cell migration assays, routine cell passage, production scale-up (bioreactor), and flow cytometry.

Trypsin / EDTA (1X solution)0.05% Trypsin in PBS with 0.1% EDTA, w/o Ca2+, Mg2+ without Phenol redTrypsin / EDTA is used for cell dissociation and for hydrolytic degradation of proteins and peptides, and is a commonly used reagent for detachment of cells from surfaces.

Supplied as a 1X ready-to-use solution.

Activity: 1g of Trypsin digests 250g of Casein substrate.

Origin: pig


GTC01.0100 Accutase® 100mL


GTC02.0100 Trypsin (1X solution) 100mL

✓ No wash

✓ No Neutralization

✓ Excellent Cell Viability

✓ Gentle

Cell Biology / Cell Detachment


www.grisp.pt

75

L-Glutamine200mM (100X)L-Glutamine is a serum-free medium supplement, useful for Biomanu- facturing, Tissue Engineering, and Specialty Media.

L-glutamine is an essential amino acid required in cell culture media formulations. Most commercially available media are formulated with free L-glutamine which is either included in the basal formula or added to liquid formulations at time of use. L-glutamine is unstable at physiologi-cal pH in liquid media. It breaks down to ammonium and pyro- glutamate at rates that make it a problem in many biomanufacturing applications.

Stable L-Glutamine200mM (100X)Stable L-Glutamine is a stabilized form of L-Glutamine (L-alanyl-L- glutamine).

As L-Glutamine is very unstable in liquid media, breaking down and releasing cytotoxic compounds, this stable form of L-Glutamine reduces cytotoxicity, by preventing degradation.

Replacing the L-glutamine in medium with stable glutamine (L-Alanyl- L-Glutamine) shows an improved stability compared to free L-Glutamine and considerable reduction of spontaneous ammonia generation. An additional benefit using stable Glutamine instead of L-glutamine in cell culture is the extension of culture time, potentially reducing the number of times the cells must be passaged.

Penicillin-Streptomycin (100X)

Penicillin-Streptomycin is a broad band antibiotic, effective against Gram (+) and Gram (–) bacteria.

Streptomycin inhibits protein synthesis, whereas penicillin inhibits cell wall synthesis.


GTC05.0100 Penicillin-Streptomycin (100x) 100mL


GTC04.0100 Stable L-Glutamine (100x) 100mL


GTC03.0100 L-Glutamine (100x) 100mL

Cell Biology / Supplements

Notes

Notes

Notes

Visit our website for promotions:www.grisp.pt/promotions

facebook.com/grispresearchsolutions

Documents

Research Solutions - GRiSPgrisp.pt/docs/grisp2016.pdfanalysis, restriction enzyme digest analysis, Northern Blotting, etc. NOTE: GRiSP Research Solutions recommends using GRS Agarose