Reaction No.1Selected Life Science Products 2014

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2 ReAction Molecular biology grade

Nucleic Acid IsolationDescription Order No. Quantity CTAB – Lysis buffer BioChemica Cetyltrimethylammonium bromide (CTaB) is used to liberate and form complexes with total nucleic acids of plants or fungi; polysaccharides, phenolic compounds and other enzyme-inhibiting contaminats found in plant cells are efficiently removed in the supernatant.

A4150,1000 1 L

Lysozyme BioChemica A3711,0050 50 gProteinase K* A3830,0500 500 mgProteinase K - Solution A4392,0010 10 mlCesium chloride 99.999 % Molecular biology grade A1098,0500 500 g1-Bromo-3-chloropropane BioChemica Chloroform substitute in phenolic DNa extraction. Bromo chloropropane is less toxic than chloroform and forms a tighter interphase!

A2107,0250 250 ml

Chloroform BioChemica A3691,0500 500 mlIsoamyl alcohol Molecular biology grade A2610,0500 500 mlTRItidy G™ Ready-to-use monophasic reagent (contains phenol and guanidinium thiocyanate) for subsequent isolation of RNa, DNa and proteins from samples of human, animal, plant and bacterial origin.

A4051,0200 200 ml

RNase A (Dnase-free) A3832,0250 250 mgPhenol equilibrated, stabilized A1153,0250 250 mlPhenol equilibrated, stabilized: Chloroform:Isoamyl alcohol 25 : 24 : 1 For DNa isolation

A0889,0250 250 ml

DNA – Isolation reagent for genomic DNA Non-organic and ready-to-use reagent for the isolation of genomic DNa from human, animal (incl. mouse tail), plant, yeast, bacterial and viral origin. The isolated DNa can be used, without additional purification, for Southern-analysis, dot blot hybridization, molecular cloning, RFLP, PCR and other molecular biology and biotechnology applications.

A3418,0050 50 ml

DEPC BioChemica A0881,0020 20 ml

DNase I A3778,0050 A3778,0500

50 mg 500 mg

Phenol water-saturated, stabilized A1624,0250 250 mlPhenol stabilized: Chloroform:Isoamyl alcohol 25 : 24 : 1 A2279,0250 250 mlRNAtidy G Ready-to-use solution for the isolation of small and large RNa species (0.1 – 15 kb) from biological material with high purity (DNa and protein-free) . Mono-phasic reagent containing phenol and guanidinium thiocyanate.

A2867,0200 200 ml

RNase-ExitusPlus™ RNase decontamination solution; removes RNase contaminations from surfaces by non-enzymatic degradation.

A7153,0250 A7153,0500

250 ml 500 ml

In the center of molecular biology is one species of molecules: DNa. DNa molecules are amplified and introduced into organisms by transformation or transfection, separated, stained, examined under the microscope, manipulated, sequenced and so on. For all these techniques the initial step is to isolate DNa from the origin of interest.

* Proteinase K When it comes to protein degradation, proteinase K is the most common protease directly employed in the lysis solution. Conditions that promote enzyme inactivation like detergents, chaotropic salts, high temperature and changes in pH are well tolerated by proteinase K. Beside its high stability, proteinase K is characterized by a large number of cleavage sites and therefore perfectly suited to remove cellular and nuclear proteins that are attached to the DNA. Furthermore, the permissive Proteinase K has no need for cofactors, so it can’t be inhibited by EDTA.

DNA contamination of PCR workstations leads to faulty DNA amplifications. Hidden traces of DNA may cause false diagnoses or contamination of sequence databases. Control and elimination of unwanted DNA background therefore is mandatory for quality control in all PCR laboratories.

Our patented product DNa-ExitusPlus™ employs a mild and non-corrosive chemistry for rapid non-enzymatic degradation of nucleic acids. Even a short incubation time with DNa-ExitusPlus™ completely removes non-target DNa and RNa from work surfaces and tools (1, 2).

AppliChem’s Solution!Nucleic Acid Decontamination with DNA-ExitusPlus™

There are two different versions of DNa-Exitus Plus™ available: DNa-ExitusPlus™ (a7089) includes a color indicator to easily visualize the surface area covered by the reagent. DNa-ExitusPlus™ IF (a7409) is almost colorless.

DNA-ExitusPlus™ is a registered trademark of AppliChem GmbH.

DNA-free Reagents for PCRDescription Order No. QuantityWater tested PCR,DNA-free a8510,1017 10 x 1,7 ml

SYBR Green®-staining solution, DNA-free a8511,50625 5 x 0,625 ml

Products for Nucleic Acid DecontaminationDescription Order No. QuantityDNA-ExitusPlus™ a7089,0100 100 ml

DNA-ExitusPlus™ a7089,0500 500 ml

DNA-ExitusPlus™ IF a7409,0100 100 ml

DNA-ExitusPlus™ IF a7409,0500 500 ml

Autoclave-ExitusPlus™ a7600,1000 Powder for 6 x 1 L

4 ReAction Molecular biology grade

It‘s harder than it seems to test decontamination solutions for a complete DNa degrading activity. It is not sufficient at all to just mix decontamination solution and DNa and to perform a subse-quent PCR to amplify residual DNa. Since PCR is too easily affected by factors such as unfavorab-le buffer conditions, PCR inhibitors or DNa cross contamination. But for the evaluation of the potential of a DNa decontamination reagent, one has to use PCR analysis in combination with a sensitive DNa degradation test. We have developed a DNa strand break assay which allows us to evaluate the efficiency of DNa-ExitusPlus™ [1].

Results While the control treatment and all tested competitors show no DNa degradation, plasmid DNa is degraded completely after 10 min of treatment with DNa-ExitusPlus™. 3 min of DNa-ExitusPlus™-treatment leads already to a strong fragmentation of the test plasmids.

Results after incubation with DNa-ExitusPlus no PCR amplicons for the test DNa can be detected indicating all test DNa template has been degraded. The PCR product for control DNa confirms functionality of the PCR.

Literature[1] Esser et al. (2006) DNA Decontamination: Novel DNA-ExitusPlus™ in comparison with conventional reagents.

BioTechniques 40 (2), 238-9. – On closer inspection, only DNA-ExitusPlus™ destroys DNA completely.[2] Arena, A. (2010) Dna Exitus Plus™ Versus Standard Bleach Solution for the Removal of Dna Contaminants on Work Surfaces and Tools. Investigative

Science Journal 2, 20-9. – DNA-ExitusPlus ™ proved as effective as freshly prepared bleach solution in an assay using swap tests and subsequent qPCR.

Please see our website or refer to our AppliCation No.1 for more details of the procedure.

Verify the effectiveness of decontamination agents

A. The DNA strand break assay in brief: 1. Incubate for 3-10 min 200 ng of plasmid DNa with

water (C), commercial decontamination solutions (X1-X4) and DNa-ExitusPlus™ (D).

2. Denature all samples with heat. 3. agarose gel electrophoresis of DNa and

degradation products.

B. The PCR test in brief: 1. Incubate 0.1 to 1 ng of test DNa with water (a)

or DNa-ExitusPlus™ (B). 2. Wash twice the reactions. 3. add a control DNa, and 4. Run PCR with primers against test and control DNa. 5. agarose gel electrophoresis of PCR products.

Agarose gel electrophoresis ReAction 5

Description Order No. QuantityDNA-Dye NonTox Non-hazardous, non-mutagenic fluorescent dye (ethidium bromide substitute) for staining of DNa in agarose gels. Detection under blue light or UV light. Supplied as a ready-to-use 6X loading dye.

a9555,1000 1 ml

Ethidium bromide - Solution 1 % BioChemica a1152,0025 25 ml

Ethidium bromide - Solution 0.07 % „dropper-bottle“ a2273,0015 15 ml

Decontamination Bag Tear-resistant charcoal bags for removal of DNa dyes such as ethidium bromide, SYBR Green® or propidium iodide from aqueous solutions.

a9676,0025 25 Stk.

Loading buffer DNA I Contains bromophenol blue and Ficoll® 400

a3144,0010 10 ml

Loading buffer DNA II Contains bromophenol blue, xylene cyanol FF and Ficoll® 400

a2571,0025 25 ml

Loading buffer DNA IV (for agarose gels) Contains bromocresol green, xylene cyanol, EDTa, SDS and Ficoll® 400

a3481,0010 10 ml

Agarose Basic a8963,1000 1 kg

Agarose high EEO a2115,0500 500 g

Agarose low EEO (agarose Standard) a2114,0500 500 g

Agarose Low Melt Large DNA grade For analytic and preparative DNa gels

a3762,0025 25 g

Agarose medium EEO a2116,0100 100 g

Agarose MP Multi-purpose agarose; very good analytic separation (100 bp to 50 kb!); blotting; DNa typing; PFGE.

a1091,0500 500 g

DNA Ladder 1 kb a5207,0005 50 µg

DNA Ladder 100 bp a5191,0005 50 µg

DNA Ladder 100 bp (lyophilised) a3470,0050 50 µg

DNA Ladder 50 bp a8368,0050 50 µg

DNA Ladder Mix 100 – 5000 (lyophilised) a3660,0050 50 µg

Ethidium bromide „dropper-bottle“ A2273 One drop of the solution stains an agarose gel of 50 ml. The dropper-bottle is convenient and minimizes the possible contact with ethidium bromide.

DNA ladder 1 kb (A3470) The mass of every band is defined and may be used for semi-quanti-tative determination of DNa concentration.

Decontamination Bags (A9676) in use Just add a „tea bag“ to the ethidium bromide-containing buffer and wait... activated charcoal will absorb the dye.

B. The PCR test in brief: 1. Incubate 0.1 to 1 ng of test DNa with water (a)

or DNa-ExitusPlus™ (B). 2. Wash twice the reactions. 3. add a control DNa, and 4. Run PCR with primers against test and control DNa. 5. agarose gel electrophoresis of PCR products.

6 ReAction Biochemistry

Enzyme substratesDescription Order No. Quantity

2-Nitrophenyl-ß-D-galactopyranoside BioChemica a1272,0025 25 g

4-Nitrophenyl phosphate disodium salt hexahydrate BioChemica a1442,0025 25 g

NADPH tetrasodium salt a1395,0001 1 g

NADP sodium salt a1394,0001 1 g

NAD a1124,0005 5 g

L-Glutathione oxidize BioChemica a2243,0025 25 g

L-Glutathione reduced BioChemica a2084,0005 5 g

ß-Glycerol phosphate disodium salt pentahydrate BioChemica a2253,0500 500 g

Denaturating, reducing, complexingDescription Order No. Quantity

Guanidine thiocyanate-Solution (6 M in 0.1 M Tris; pH 7.5) Molecular biology grade a0703,1000 1 L

Guanidine thiocyanate Molecular biology grade a1107,0500 500 g

Guanidine hydrochloride Molecular biology grade a1106,0500 500 g

Urea Molecular biology grade a1049,1000 1 kg

DTE BioChemica a1102,0025 25 g

DTT BioChemica a1101,0100 100 g

DTT - Solution (1 M) Molecular biology grade a3668,0050 50 ml

ß-Mercaptoethanol Molecular biology grade a1108,0100 100 ml

EDTA BioChemica a1103,1000 1 kg

EDTA disodium salt dihydrate BioChemica a1104,1000 1 kg

EDTA Molecular biology grade a5097,0500 500 g

EDTA-Solution pH 8.0 (0.5 M) Molecular biology grade a4892,0500 500 ml

EGTA für die Molekularbiologie a0878,0025 25 g

IPTG Molecular Biology grade a4773,0025 25 g

X-Gal Molecular Biology gradeA4978,0001 1 g

example

Gel Filtration ReAction 7

appliChem Panreac offers ready-to-use gravity flow and spin columns, and columns for FPLC systems, all products prepacked with appli-Xchange. appliXchange is a white solid com-posed of polymerized and cross-linked dextran: The beaded composite material is prepared in water or any desired aqueous buffer solution and subsequently cast into suitable columns. Molecules purified with appliXchange are se-parated according to size. Smaller molecules pass significantly slower through the column than larger molecules. This is due to the longer path the smaller molecules must travel. This longer path arises from the pores of the beads. Due to the multitude of pores, most of the total volume of the beads is in fact water. Small mole cules enter the pores and get into the beads. The delay caused by entering the pores corre-

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Elution volume [mL]0,0 1,0 2,0 3,0 4,0 5,0 6,0 7,0

Elution profile for removal of excess FITC from IgG after coupling reaction. 1 mg IgG anti-Rabbit and 0.1 µmol FITC in 1 ml DMSO/NaHCO3; IgG (280 nm): black line; FITC (490 nm): magenta line.

lates with the molecular size. Molecules larger than the size exclusion cut-off (also referred to as molecular weight cut-off or size exclusion limit) cannot enter the pores and therefore flow around the porous beads rapidly passing the column.

Size Exclusion Chromatography (SEC, also referred to as Gel Filtration) is a chromatographic purification method for size-based separation of molecules in solution. Commonly this technique is used for large water soluble bio-molecules, such as DNA, oligonucleotides, proteins, antibodies and other polymers.

Rapid Purification of Bio-Molecules

8 ReAction Gel Filtration

What is the difference between G50 and G25?The number in the product description correlates to the swelling power of the matrix: Grade50 indicates that the dry matrix material will take up 5.0 x times its weight of water. G25 absorbs 2.5 x times its weight of water. The higher the swelling power the larger the pore size. The pore size determines the size exclusion cut-off.

What does „cut-off: 5 kD/10 bp“ mean?The cut-off gives the maximum molecule size that will be retained by the column. a size ex-clusion cut-off of 5 kDa means, that molecules of a smaller size will remain (a certain time) in the matrix pores, while larger molecules rapidly run through the matrix.

Why offering different particle sizes?The particle size or bead size does not influ-ence the separation range or cut-off characte-ristics of the matrix, but is correlating to the flow rate. Small particles make the matrix more stable and are well suitable for centrifugation or vacuum. On the other hand, larger particle sizes enable adequate flow rates when using gravity based columns.

The SEC matrix by scanning electron microscopy.

AppliXchange G25-SF particles on a graphite surface.

FAQs and Important Information

Buffer conditions, pH value and temperature during the filtration process may be modi-fied according to the needs of the molecules but not the column material. SEC can be per-formed in the presence of detergents, urea (up to 8 M) and guanidine hydrochloride (6 M), organic solvents (e.g. 24 % ethanol; 30 % propanol; 30 % acetonitrile), co-factors or ions, under basic and acidic conditions, high or low ionic strength.

Hydrated gel filtration column for rapid and efficient removal of small molecules (salts, dyes, ammonia, haptens, biotin, etc.) from proteins, nucleic acids and

other macromolecules. DextraSEC PRO (for protein purification) and DextraSEC NA (for nucleic acids) are available for processing sample sizes from 200 µl to 50 ml.

Ready-to-Use Gel Filtration Columns and PlatesDextraSEC and DextraSEP are ready-to-use gel filtration columns available in many diffe-rent sizes (to process sample volumes from 10 µl up to 50 ml) and formats (gravity columns, spin columns, multiwell plates, FPLC columns) for convenient desalting and buffer exchange.

Column matrix: appliXchange G25 M Cut off: > 5 kDa;10 bpBead size (dry): 50 – 150 µm

Gravity flow columns

FPLC columnsColumn matrix: appliXchange G25 SF Cut off: > 5 kDa; 10 bpBead size (dry): 20 – 50 µmFlow rate: 1 to 10 ml/min. Max. Backpressure: 3 bar

Hydrated gel filtration column for desalting, separa tion of larger bio-molecules (i.e. proteins such as anti bodies, enzymes or larger nucleic acids), and buffer exchange

using Liquid Chromatography system. DextraSEP FPLC1 (a9790) and DextraSEP FPLC5 (a9749) can take maximum sample volumes of 0.3 and 1.5 ml.

NEW

Please visit our website www.applichem.com

Hydrated gel filtration multi-well plates for high-throughput applications (e.g. removal of excess Dye terminators from completed DNA sequencing

reactions). DextraSEC 96W, DextraSEC 96W-large, and DextraSEC 384W are designed for sample volumes 15, 40, and 10 µl, respectively.

Column matrix: appliXchange G50 SF Cut off: > 25 kDa; 20 bpBead size (dry): 20 – 50 µm

Multiwell plates

10 ReAction Gel Filtration

Description Order No. Content

DextraSEC Mini-spinPRO Desalt (G-25)Purified proteins are eluted into pure water a9724,0004 4 Columns

DextraSEC Mini-spinPRO Desalt (G-25), stabilized Purified Proteins are eluted into stabilized water a9708,0004 4 Columns

DextraSEC Mini-spinPRO PBS (G-25) Elution into PBS, pH 7 a8566,0004 4 Columns

DextraSEC Mini-spinPRO TRIS (G-25) Elution into pH 6 Tris buffer solution a9692,0004 4 Columns

DextraSEC Mini-spinNA Desalt (G-25)Purified nucleic acids are eluted into pure water a9700,0004 4 Columns

Mini-spin columns for rapid desalting or buffer exchange

Description Order No. Content

DextraSEC Mini-spinPRO Desalt (G-50) Purified proteins are eluted into pure water a9776,0004 4 Columns

DextraSEC Mini-spinPROPBS (G-50) Elution into PBS, pH 7 a9763,0004 4 Columns

DextraSEC Mini-spinPRO TRIS (G-50) Elution into Tris, pH 6 a9741,0004 4 Columns

DextraSEC Mini-spinNA Desalt (G-50) Purified nucleic acids are eluted into pure water a8563,0004 4 Columns

Column matrix: appliXchange G50 SF Cut off: > 25 kDa; 20 bpBead size (dry): 20 – 50 µm

Bed volume: 0.5 ml Sample volume: 10 – 100 µl

Column matrix: appliXchange G25 SF Cut off: > 5 kDa; 10 bpBead size (dry): 20 – 50 µm

Bed volume: 0.5 ml Sample volume: 10 – 100 µl

NEWPre-packed and ready to go! Clean proteins or nucleic acids in 5 minutes only

1. Spin column and discard storage buffer2. Apply sample3. Spin to elute purified sample into your

preferred buffer solution

AntibioticsDescription Order No. Quantity

Amphotericin B BioChemica a1907,0050 50 mg

Ampicillin sodium salt BioChemica

a0839,0100 100 g

Chloramphenicol BioChemica a1806,0050 50 g

Cycloheximide BioChemica a0879,0005 5 g

Doxycycline hyclate BioChemica a2951,0025 25 g

G418 disulfate BioChemica a2167,0001 1 g

G418 disulfate – Solution, sterile a6798,0050 50 ml

Gentamycin sulfate BioChemica a1492,0005 5 g

Hygromycin B a5347,0250 250 mg

Hygromycin B - Solution a2175,0025 25 ml

Kanamycin sulfate BioChemica A1493,0010 10 g

Penicillin - Streptomycin (100X) Cell culture grade

a8943,0100 100 ml

Penicillin G potassium salt BioChemica

a1837,0025 25 g

Polymyxin B sulfate BioChemica a0890,0005 5 g

Spectinomycin dihydrochloride pentahydrate BioChemica

a3834,0005 5 g

Streptomycin sulfate BioChemica

a1852,0025 25 g

Vancomycin hydrochloride BioChemica

a1839,0001 1 g

a bit more ReAction 11

Salts & BuffersDescription Order No. Quantity

Magnesium chloride hexahydrate BioChemica

a1036,0500 500 g

Sodium azide pure a1430,0100 100 g

Sodium chloride BioChemica a1149,5000 5 kg

Sodium chloride - Solution (5 M) Molecular biology grade

a7006,1000 1 L

Sodium hydroxide pellets Molecular biology grade

a6829,1000 1 kg

Potassium chloride BioChemica a1039,1000 1 kg

PBS tablets pH 7.4 (for 1 L) a9201,0100 100 Tabs

PBS tablets pH 7.2 (for 1 L) a9202,0010 10 Tabs

PBS buffer (1X, Dulbecco‘s) - Powder

a0964,9050 50 L

Silver nitrate Molecular biology grade

a3944,0025 25 g

Silver nitrate BioChemica a3972,0100 100 g

Tris Molecular biology grade

A2264,1000 1 kg

Tris hydrochloride Molecular biology grade

a3452,0500 500 g

TAE buffer (50X) Molecular biology grade

a4686,1000 1 L

TBE buffer (10X) Molecular biology grade

a3945,1000 1 L

SolventsDescription Order No. Quantity

Aceton BioChemica a3855,2500 2,5 L

Dimethyl sulfoxide Molecular biology grade a3006,0500 500 ml

Ethanol absolute Molecular biology grade A3678,1000 1 L

Acetic acid 100 % BioChemica a3701,2500PE 2,5 L

Formaldehyde - Solution 37 % Molecular biology grade a0877,0250 250 ml

Formamid BioChemica a0937,2500 2,5 L

Glycerol 87 % BioChemica a0970,1000 1 L

Glycerol 87 % Molecular biology grade a3739,0500 500 ml

Methanol BioChemica a3493,2500PE 2,5 L

2-Propanol BioChemica a3465,2500 2,5 L

2-Propanol Molecular biology grade a3928,1000PE 1 L

Hydrochloric acid (1 M) Molecular biology grade a6578,1000 1 L

Trichloroacetic acid - Solution 20 % BioChemica a0590,0500 500 ml

Water bidistilled, sterilea4042,0500 a4042,1000

500 ml 1 L

For more information and more Life Science products please visit

www.applichem.com