Product Class: Other

EpiMark® Hot Start Taq DNA Polymerase
NEBU cloned at NEB recombinant No hotstart pcr 5 Epi

Product Introduction

Taq DNA Polymerase optimized for epigenetics analysis

  • Ideal for use on bisulfite-converted DNA and AT-rich templates
  • Specially formulated reaction buffer system
  • Aptamer-based hot start technology avoids nonspecific amplification and enables room temperature reaction setup
Catalog # Size Concentration
M0490S 100.0 reactions 5000 units/ml
M0490L 500.0 reactions 5000 units/ml

Product Information

Description

EpiMark Hot Start Taq DNA Polymerase is a mixture of Taq DNA Polymerase and a temperature sensitive, aptamer-based inhibitor. The inhibitor binds reversibly to the enzyme, inhibiting polymerase activity at temperatures below 45°C, but releases the enzyme during normal PCR cycling conditions. This permits assembly of reactions at room temperature. An advantage of the aptamer-based hot start mechanism is that it does not require a separate high temperature incubation step to activate the enzyme. The advanced aptamer-based hot-start activity coupled with the supplied optimized reaction buffer makes the EpiMark Hot Start Taq DNA Polymerase an excellent choice for use on bisulfite-converted DNA.

Taq DNA polymerase possesses a 5´- 3´ polymerase activity (1,2,3) and a 5´ flap endonuclease activity (4,5).

EpiMark Hot Start Taq DNA Polymerase is supplied with 5X EpiMark Hot Start Taq Reaction Buffer, providing robust amplification for bisulfite-converted DNA and AT-rich amplicons.

Product Source

An E. coli strain that carries the Taq DNA Polymerase gene from Thermus aquaticus YT-1
This product is related to the following categories:
Methylome Analysis Products,
Epigenetic Analysis Products,
Taq DNA Polymerase Products,
Next Generation Sequencing Library Preparation Products,
This product can be used in the following applications:
Bisulfite Sequencing,
Bisulfite Conversion,
Hot Start PCR,
Specialty PCR,
Routine PCR,
PCR

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • M0490S     -20    
      EpiMark® Hot Start Taq DNA Polymerase M0490SVIAL -20 1 x 0.025 ml 5,000 units/ml
      EpiMark® Hot Start Taq Reaction Buffer Pack B0490SVIAL -20 1 x 2 ml 5 X
  • M0490L     -20    
      EpiMark® Hot Start Taq DNA Polymerase M0490LVIAL -20 1 x 0.125 ml 5,000 units/ml
      EpiMark® Hot Start Taq Reaction Buffer Pack B0490SVIAL -20 3 x 2 ml 5 X

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme that will incorporate 15 nmol dNTP into acid insoluble material in 30 minutes at 75°C.

Reaction Conditions

1X EpiMark® Hot Start Taq Reaction Buffer Pack

1X EpiMark® Hot Start Taq Reaction Buffer Pack
20 mM Tris-HCl
1.8 mM MgCl2
22 mM KCl
0.06% IGEPAL® CA-630
0.05% Tween® 20
22 mM NH4Cl
(pH 8.9 @ 25°C)

Storage Buffer

10 mM Tris-HCl
100 mM KCl
1 mM DTT
0.1 mM EDTA
0.5% Tween® 20
0.5% IGEPAL® CA-630
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

No

Molecular Weight

Theoretical: 94000 daltons

5' - 3' Exonuclease

Yes

3' - 5' Exonuclease

No

Resulting Ends

  • Single-base 3´ Overhangs

Unit Assay Conditions

1X ThermoPol® Reaction Buffer, 200 µM dNTPs including [3H]-dTTP and 15 nM primed M13 DNA. 

Application Features

  • PCR from bisulfite-converted DNA

References

  1. Chien, A., Edgar, D.B. and Trela, J.M. (1976). J. Bact. . 127, 1550-1557.
  2. Kaledin, A.S., Slyusarenko, A.G. and Gorodetskii, S.I. (1980). Biokhimiya . 45, 644-651.
  3. Lawyer, F.C. et al. (1993). PCR Methods And Appl.. 2, 275-287.
  4. Longley, M.J., Bennett, S.E. and Mosbaugh D.W. (1990). Nucleic Acids Res. . 18, 7317-7322.
  5. Lyamichev, V., Brow, M.A. and Dahlberg, J.E. (1993). Science. 260, 778-783.
  6. Saiki R.K. et al. (1985). Science. 230, 1350-1354.
  7. Powell, L.M. et al. (1987). Cell. 50, 831-840.
  8. Sun, Y., Hegamyer, G. and Colburn, N. (1993). Biotechniques . 15, 372-374.

Protocols, Manuals & Usage

Protocols

  1. EpiMark® Hot Start Taq DNA Polymerase Guidelines for PCR (M0490)

Usage & Guidelines

Tools & Resources

Web Tools

FAQs & Troubleshooting

FAQs

  1. What ends will my PCR products have?
  2. What are Hot Start and WarmStart® polymerases and when would I use them?
  3. How should I determine the appropriate annealing temperature for my reaction?
  4. My results are not as expected. Where can I find troubleshooting help?
  5. What should the final primer concentration be in my PCR?
  6. Is this Taq DNA Polymerase product compatible with other NEB Buffers?
  7. What are the general recommendations for designing primers for bisulfite-treated/deaminated DNA?

Quality, Safety & Legal

Quality Assurance Statement

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

Specifications

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

Certificate Of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

Safety DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email [email protected].

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

Licenses

Nucleic acid-based aptamers for use with thermophilic DNA polymerases are licensed exclusively by New England Biolabs, Inc. from SomaLogic, Inc. New England Biolabs, Inc. gives the Buyer/User a non-exclusive license to use EpiMark Hot Start Taq DNA Polymerase for Research Use Only (RUO). Commercial use of this product may require a license from New England Biolabs, Inc. For additional information or to inquire about commercial use, please contact [email protected]. 

Trademarks

NEW ENGLAND BIOLABS®, EPIMARK® and THERMOPOL® are registered trademarks of New England Biolabs, Inc.
IGEPAL® is a registered trademark of Rhodia  Operations.
TWEEN® is a registered  trademark of Uniqema americas LLC.

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