OneTaq DNA Polymerase - McGill University · OneTaq DNA Polymerase versus two leading non-hot start “enhanced” Taq polymerases. The yield and the % purity of the reaction products

PRODUCT NEB# LIST PRICE PROMOTIONAL PRICE *

OneTaq® DNA Polymerase

M0480S - 200 units $58 $40.60

M0480L - 1,000 units $233 $163.10

M0480X - 5,000 units $914 $639.80

OneTaq® Hot Start DNA Polymerase

M0481S - 200 units $121 $84.70

M0481L - 1,000 units $483 $338.10

M0481X - 5,000 units $1,931 $1351.70

OneTaq ® 2X Master Mix with Standard BufferM0482S - 100 rxns $58 $40.60

M0482L - 500 rxns $256 $179.20

OneTaq ® 2X Master Mix with GC BufferM0483S -100 rxns $58 $40.60

M0483L - 500 rxns $256 $179.20

OneTaq® Hot Start 2X Master Mix with Standard BufferM0484S - 100 rxns $98 $68.60

M0484L - 500 rxns $443 $310.10

OneTaq ® Hot Start 2X Master Mix with GC BufferM0485S - 100 rxns $98 $68.60

M0485L - 500 rxns $443 $310.10

OneTaq ® Quick-Load 2X Master Mix with Standard BufferM0486S - 100 rxns $58 $40.60

M0486L - 500 rxns $256 $179.20

OneTaq ® Quick-Load 2X Master Mix with GC BufferM0487S - 100 rxns $58 $40.60

M0487L - 500 rxns $256 $179.20

OneTaq ® Hot Start Quick-Load 2X Master Mix with Standard BufferM0488S - 100 rxns $98 $68.60

M0488L - 500 rxns $443 $310.10

OneTaq ® Hot Start Quick-Load 2X Master Mix with GC BufferM0489S - 100 rxns $98 $68.60

M0489L - 500 rxns $443 $310.10

Advantages of OneTaq® DNA Polymerase Enzymes• Blend of Taq and Deep Vent, optimized for

robust amplification across the broadest range of templates

• Fidelity is 2X that of Taq DNA Polymerase

• Little or no optimization is required to achieve high yields

Advantages of OneTaq’s aptamer-based hot start technology• Room temperature reaction set-up

• No separate activation incubation required

• Value pricing

• Polymerase retains high specific activity

Applications for OneTaq® • Routine PCR

• Colony PCR

• SNP Detection

• T/A, U/A Cloning

• Genotyping

The figure exemplifies the superior performance, in this case with high GC amplicons, of OneTaq DNA Polymerase versus two leading non-hot start “enhanced” Taq polymerases. The yield and the % purity of the reaction products are shown.

OneTaq DNA Polymerase is an optimized blend of Taq and Deep VentR

™ DNA polymerases for use

with routine and difficult PCR experiments. The 3´→ 5´ exonuclease activity of Deep Vent DNA Polymerase increases the fidelity and robust amplification of Taq DNA Polymerase (1). OneTaq Reaction Buffers and High GC Enhancer have been formulated for robust yields with minimal optimization, regardless of a template’s GC content.

OneTaq DNA Polymerase is supplied with two 5X buffers: (Standard and GC), as well as a High GC Enhancer solution. For most routine, AT-rich or complex amplicons with up to ~65% GC content, OneTaq Standard Reaction Buffer provides robust amplification. For GC-rich amplicons, the OneTaq GC Reaction Buffer can improve both performance and yield. For particularly high GC or difficult amplicons, the OneTaq High GC Enhancer can be added to reactions containing OneTaq GC Reaction Buffer.

In addition to standalone enzymes, both OneTaq and OneTaq Hot Start DNA Polymerases are available in master mix and Quick-Load master mix formats. Master mix formulations include dNTPs, MgCl2 and other buffers and stabilizers. The Quick-Load master mix formulations also include two tracking dyes for use with downstream visualization (i.e., agarose gels). With these convenient formats, the addition of primers and template are all that is required for robust amplification.

OneTaq® DNA Polymerase

OneTaq ® DNA Polymerase

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66% 68% 68% 79% 80%

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NEB OneTaq DNA Polymerase Competitor 1 Competitor 2

NEB OneTaq DNA PolymeraseCompetitor 1: GoTaq (Promega)Competitor 2: DreamTaq (Fermentas/Thermo)

Yiel

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*TERMS & CONDITIONS: Offer valid in Canada only. Expires April 30th, 2017. Discount is eligible for products listed on this flyer and can only be obtained through a quote or purchasing online at www.neb.ca. Promotion not valid for cash or cash equivalent towards purchase(s). No substitutions. Offer may not be applied to existing, pending or prior orders. Cannot be combined with any other promotion or discount.

One or more of these products are covered by patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc. For more information, please email us at [email protected]. The use of these products may require you to obtain additional third party intellectual property rights for certain applications.

New England Biolabs Ltd. is the exclusive Canadian distributor for Cell Signaling Technology, Inc.New England Biolabs Ltd. Tel: 1-800-387-1095 | Fax: 1-800-563-3789 | [email protected] | [email protected] | www.neb.ca

Promotion valid until April 30th, 2017 For more information visit: www.neb.ca/onetaq-promo

Julie Wong, Ph.D. Territory Manager, Western CanadaCell: (604) 767-5078 [email protected]

Howard Cukier, Ph.D. Territory Manager, OntarioCell: (416) [email protected]

Alexandre Rochette, Ph.D. Territory Manager, QuebecCell: (514) [email protected]

Western blotting ITEM#/SIZEPROMOTIONAL

PRICE*

Anti-rabbit IgG, HRP-linked Antibody

7074P2 -100 ul $74.25

7074S - 1 ml $138.75

7074V - 5 ml $441.75

Anti-mouse IgG, HRP-linked Antibody

7076P2 - 100 ul $74.25

7076S - 1 ml $138.75

7076V - 5 ml $441.75

Anti-rat IgG, HRP-linked Antibody 7077S - 1 ml $138.75

Anti-biotin, HRP-linked Antibody 7075S - 1 ml $138.75

Anti-rabbit IgG, AP-linked Antibody 7054S - 1 ml $138.75

Anti-mouse IgG, AP-linked Antibody 7056S - 1 ml $138.75

Anti-biotin, AP-linked Antibody 7055S - 1 ml $138.75

Mouse Anti-rabbit IgG (Light-Chain Specific) (L57A3) mAb 3677S - 100 ul $150.00

Mouse Anti-rabbit IgG (Conformation Spectic) (L27A9) mAb 3678S - 100 ul $150.00

Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) 5127S - 100 ul $177.75

Anti-biotin (D5A7) Rabbit mAb (HRP Conjugate) 5571S -1 ml $135.75

Anti-biotin (D5A7) Rabbit mAb 5597S - 100 ul $258.00

Anti-rabbit IgG (H+L), Biotinylated Antibody 14708S - 1 ml $146.25

Anti-mouse IgG (H+L), Biotinylated Antibody 14709S - 1 ml $146.25

Mouse Anti-rabbit IgG (Light-Chain Specific) (D4W3E) mAb 45262S - 100 ul $183.00

Anti-Blue (2D2F11) Mouse mAb (HRP Conjugate) 46387S - 100 ul $219.75

Rabbit Anti-Mouse IgG (Light Chain Specific) (D3V2A) mAb (HRP Conjugate) 58802S -100 ul $177.75

Immunohistochemical ITEM#/SIZEPROMOTIONAL

PRICE*

SignalStain® Boost IHC Detection Reagent (HRP, Rabbit)8114P - 1 ml $41.25

8114S - 15 ml $377.25

SignalStain® Boost IHC Detection Reagent (HRP, Mouse)8125P - 1 ml $41.25

8125S - 15 ml $377.25

Immunofluorescent/Flow cytometric ITEM#/SIZEPROMOTIONAL

PRICE*

Anti-mouse IgG (H+L), F(ab’)2 Fragment (Alexa Fluor® 488 Conjugate) 4408S - 250 ul $146.25



Anti-rabbit IgG (H+L), F(ab’)2 Fragment (Alexa Fluor® 488 Conjugate) 4412S - 250 ul $146.25



Anti-rat IgG (H+L), (Alexa Fluor® 488 Conjugate) 4416S - 250 ul $125.25



Anti-rabbit IgG (H+L), F(ab’)2 Fragment (PE Conjugate) 8885S - 250 ul $209.25

Anti-mouse IgG (H+L), F(ab’)2 Fragment (PE Conjugate) 8887S - 250 ul $209.25



Anti-Rabbit IgG (H+L), (PE Conjugate) 14705S - 250 ul $146.25

Secondary Antibodies From Cell Signaling Technology®

For more information visit: www.neb.ca/offers

Julie Wong, Ph.D. Territory Manager, WesternCanadaCell: (604) 767-5078 | [email protected]

Howard Cukier, Ph.D. Territory Manager, OntarioCell: (416) 527-2352 | [email protected]

Alexandre Rochette, Ph.D. Territory Manager, QuebecCell: (514) 233-4557 | [email protected]

*TERMS & CONDITIONS: Offer valid in Canada only. Expires March 31st, 2017. Discount is eligible for products listed on this flyer and can only be obtained through a quote or purchasing online at www.neb.ca. Promotion not valid for cash or cash equivalent towards purchase(s). No substitutions. Offer may not be applied to existing, pending or prior orders. Cannot be combined with any other promotion or discount.

New England Biolabs Ltd. is the exclusive Canadian distributor for Cell Signaling Technology, Inc.New England Biolabs Ltd. Tel: 1-800-387-1095 | Fax: 1-800-563-3789 [email protected] | [email protected] | www.neb.ca

For Research Use Only. Not For Use In Diagnostic Procedures.

Alexa Fluor is a registered trademark of Life Technologies Corporation.SignalStain and Cell Signaling Technology are trademarks of Cell Signaling Technology, Inc.

For your convenience, Cell Signaling Technology® (CST) offers a wide selection of secondary antibodies, chemical modulators, and detection reagents, as well as standard buffers and experimental controls. These same reagents are also used in-house for antibody validation in applications including western blotting, immunohistochemistry, flow cytometry, and immunofluorescent analysis and therefore work optimally with our primary antibodies.

Genome Editing: Tools for CRISPR/Cas9 Applications

Featured NEB Products Supporting CRISPR WorkflowsNEB # PRODUCT NAME CRISPR/CAS9 APPLICATION

M0646T/M EnGen Cas9 Nuclease NLS, S. pyogenes

in vitro cleavage of dsDNA. Genome engineering by direct introduction of active nuclease complexes

E3321S EnGen Mutation Detection Kit Determination of the targeting efficiency of genome editing protocols

E3322S EnGen sgRNA Synthesis Kit Generation of microgram quantities of custom sgRNA in under 1h

M0386S/T/M Cas9 Nuclease, S. pyogenes in vitro cleavage of dsDNA. Genome engineering by direct introduction of active nuclease complexes

E0554S/E0552S Q5 Site-directed Mutagenesis Kit (with or without competent cells)

Insertion of target sequence into the Cas9-sgRNA construct and modification of HDR templates

M0494S/L Q5 High-fidelity DNA Polymerases High-fidelity construct generation for use with CRISPR workflows

E2621S/L/X NEBuilder HiFi DNA Assembly Master Mix

Single-tube, isothermal generation of the Cas9-sgRNA construct and HDR templates

E5520S NEBuilder HiFi DNA Assembly Cloning Kit

Single-tube, isothermal generation of the Cas9-sgRNA construct and HDR templates (comes with competent cells)

E2060S/E2065S HiScribe T7 ARCA mRNA Kit (with or without tailing) Generation of Cas9 mRNA with ARCA cap

E2050S HiScribe T7 Quick High Yield RNA Synthesis Kit

Generation of sgRNA and Cas9 Concentration of up to 10mg/ml in under 2 hours

M0302S/L T7 Endonuclease I Determination of the targeting efficiency of genome editing protocols by surveyor assay

New England Biolabs Ltd. is the exclusive Canadian distributor for Cell Signaling Technology, Inc.New England Biolabs Ltd. Telephone: 1-800-387-1095 | Fax: 1-800-563-3789 | [email protected] | [email protected] | www.neb.ca

Genome editing is enabled by the development of tools to make precise, targeted changes to the genome of living cells. Recently a new tool based on a bacterial CRISPR-associated protein-9 nuclease (Cas9) from Streptococcus pyogenes has generated considerable excitement. This follows several attempts over the years to manipulate gene function, including homologous recombination and RNA interference. RNAi, in particular, became a laboratory staple enabling inexpensive and high-throughput interrogation of gene function, but is hampered by providing only temporary inhibition of gene function and unpredictable off-target effects.

The simplicity of the CRISPR nuclease system, with only three components (Cas9 , crRNA and trRNA) makes this system amenable to adaptation for genome editing. By combining the crRNA and trRNA into a single synthetic guide RNA (sgRNA), a further simplified two component system can be used to introduce a targeted double stranded break.

*TERMS & CONDITIONS: Offer valid in Canada only. Expires March 31st, 2017. Discount is eligible for products listed on this flyer and can only be obtained through a quote or purchasing online at www.neb.ca only for products listed on this flyer. Promo code “NEBCRISPR“ must be entered at time of online purchase. A promo code box will appear in “View Cart” page. Promotion not valid for cash or cash equivalent towards purchase(s). No substitutions. Offer may not be applied to existing, pending or prior orders. Cannot be combined with any other promotion or discount.One or more of these products are covered by patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc. For more information, please email us at [email protected]. The use of these products may require you to obtain additional third party intellectual property rights for certain applications.

Promo Code: NEBCRISPR

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Cas9 and sgRNA targeting a human gene were delivered to HEK293 cells by transfection. 2) Transfected plasmid DNA contained expression cassettes for 2x NLS (N- and C-terminal) Cas9 and sgRNA (delivered using TransIT-X2 (Mirus)). 3) Transfected mRNA was modified with pseudouridine and 5-methylcytosine and encoded 2x NLS (Nand C-terminal) Cas9. sgRNA was co-transfected with the mRNA using TransIT-mRNA. 4) Cas9 RNPs were delivered in reverse transfections using Lipofectamine RNAiMAX (Life Technologies) using 10 nanomolar final concentration of ribonucleoprotein (RNP). Cas9 has no NLS in the protein sequence. 5) EnGen Cas9 has N- and C-terminal NLSs. The efficiency of editing was determined using T7 Endonuclease I assay and is expressed as % modification.

Monarch DNA Cleanup Kits, available for both gel extraction and reaction cleanup, are optimized for maximum performance and minimal environmental impact. These kits rapidly and reliably purify up to 5 μg of concentrated, high-quality DNA and utilize a bind/wash/elute workflow with minimal incubation and spin times. The columns provided with each kit are specifically designed to eliminate buffer retention and the risk of carryover of contaminants, enabling elution of sample in volumes as low as 6 μl. Optimized buffer systems eliminate the need for monitoring and adjusting pH.

The Monarch™

DNA Gel Extraction Kit can be used to quickly purify DNA from agarose gels. Unlike other kits, there is no need to add isopropanol to the melted agarose prior to loading on the column, saving you a step.

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PRODUCT NEB # LIST PRICE PROMOTIONAL PRICE

Monarch DNA Gel Extraction Kit T1020S - 50 preps $121 $84.70

T1020L - 250 preps $560 $392.00

Monarch PCR & DNA Cleanup Kit (5 µg) T1030S -50 preps $121 $84.70

T1030L - 250 preps $560 $392.00

Ordering Information

Benefits• Elute in as little as 6 µl

• Eliminate buffer retention and salt carryover with optimized column design

• Columns and buffers available separately

• Reduce processing time with faster protocols

• Monitor completion of certain steps and easily identify buffers with colored buffer system

• No need to add RNase before starting

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Request a free sample from your Territory Manager

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Monarch™ DNA Cleanup Kits

Specifications• Binding Capacity: up to 5 µg

• DNA Size Range: ~50 bp to 25 kb

• Typical Recovery: DNA (50 bp to 10 kb): 70–90% DNA (11–25 kb): 50–70%

• Elution Volume: ≥ 6 µl

• Purity: A260

/A280

≥ 1.8

• Protocol Time: Gel Extraction: 10 min of spin and incubation time PCR & DNA Cleanup: 5 min of spin and incubation time

• Compatible Downstream Applications: ligation, restriction digestion, labeling and other enzymatic manipulations, library construction and DNA sequencing

Preps were performed according to recommended protocols. 1 µg of a 3 kb DNA fragment was incubated with 1 µM primers and OneTaq® Quick-Load® 2X Master Mix (NEB #M0486). DNA was eluted in 20 µl (NEB) and 40 µl (Qiagen) Elution Buffer. Half of the total elution volume was digested with 5 units of DraIII-HF® (NEB #R3510). The digest and the unused portion of the elution were resolved on a 1% w/v agarose gel along with a representative sample of the starting material.

Monarch PCR & DNA Cleanup Kit (5 µg) performs equivalently to the leading supplier

A B C A B CA B C A B C

NEB Qiagen

A – Column loadB – Column elutionC – DraIII digest

One microgram aliquots of a 3 kb fragment were resolved on a 1% w/v agarose gel, excised, and processed with different kits using manufacturer-specified minimum elution volumes. Values reported are the concentration and purity data determined by Nanodrop™ readings, as well as recovery calculations based on the eluted DNA concentration and recovered volume.

DNA purified using the Monarch DNA Gel Extraction Kit is recovered with similar efficiency and purity as the leading supplier, but is more highly concentrated, facilitating its use in downstream applications

Monarch QIAquick® MinElute®

1.9+/-0.08 1.9+/-0.08 1.9+/-0.04

0.80+/-0.05 0.72+/-0.09 0.66+/-0.13

144.9+/-9.6 25.7+/-3.1 73.8+/-14.2

A260/280

Yield (µg)

Conc. (ng/µl)

Column tip is compatible with vacuum manifolds

Made with less plastic for reduced environmental impact

Unique, tapered design eliminates buffer

carryover & allows for elution in as little as 6 μl

The improved design of the columns supplied with the Monarch DNA Gel Extraction and PCR & DNA Cleanup Kits enables elution in as little as 6 µl, and eliminates buffer retention

NEB Qiagen®

Buffer retention

Monarch columns are designed without a frit, which eliminates buffer retention and the risk of carryover contamination, providing fast, worry-free DNA purification.





*TERMS & CONDITIONS: Offer valid in Canada only. Expires March 31st, 2017. Discount is eligible for products listed on this flyer and can only be obtained through a quote or purchasing online at www.neb.ca only for products listed on this flyer. Promotion not valid for cash or cash equivalent towards purchase(s). No substitutions. Offer may not be applied to existing, pending or prior orders. Cannot be combined with any other promotion or discount.

One or more of these products are covered by patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc. For more information, please email us at [email protected]. The use of these products may require you to obtain additional third party intellectual property rights for certain applications.

NEW ENGLAND BIOLABS®, NEB® and HF® are registered trademarks of New England Biolabs, Inc.MONARCH™ is a trademark of New England Biolabs, Inc.QIAGEN®, QIAQUICK®, QIAPREP® and MINELUTE® are registered trademarks of Qiagen Group.LIPOFECTAMINE™ and NANODROP™ are trademarks of Thermo Fisher Scientific.

200

175

150

125

100

75

50

25

0Monarch QIAquick MinElute

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rage

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ratio

n (n

g/µ

l)

Order online visit: www.neb.ca/monarchpromo

Documents

OneTaq DNA Polymerase - McGill University · OneTaq DNA Polymerase versus two leading non-hot start “enhanced” Taq polymerases. The yield and the % purity of the reaction products