OneTaq® Hot Start Quick-Load® 2X Master Mix with Standard Buffer

Description

OneTaq® Hot Start Quick-Load® 2X Master Mix with Standard Buffer is an optimized, ready-to-use blend of Taq and Deep VentR™ DNA Polymerases combined with an aptamer-based inhibitor. This enzyme blend is ideally suited to routine PCR applications from a variety of templates, including pure DNA solutions, bacterial colonies and cDNA products. The 3´→5´ exonuclease activity of Deep Vent DNA Polymerase increases the fidelity and robust amplification of Taq DNA Polymerase (1). The hot start nature of the enzyme offers convenience with decreased interference from primer dimers and secondary products. The convenient Quick-Load master mix formulation contains dNTPs, MgCl2, buffer components and stabilizers as well as two commonly used tracking dyes for DNA gels. On a 1% agarose gel in 1X TBE, Xylene Cyanol FF migrates at ~4 kb and Tartrazine migrates at ~10 bp. Both dyes are present in concentrations that do not mask co-migrating DNA bands.

Amplification of a selection of sequences with varying GC content from human and C. elegans genomic DNA using OneTaq Hot Start DNA Polymerase. GC content is indicated below gel. Marker M is the 1 kb DNA Ladder (NEB #N3232 ).
  

Product Source

An E. coli strain that carries the Taq DNA Polymerase gene from Thermus aquaticus YT-1 and an E. coli strain that carries the Deep VentR DNA Polymerase gene from Pyrococcus species GB-D.

Advantages and Features

Applications

  • High Sensitivity PCR
  • High Throughput PCR
  • Routine PCR
  • AT-rich PCR
  • Colony PCR
  • Long PCR (up to ~6 kb genomic)

Properties and Usage

Storage Temperature

-20°C

1X Master Mix Composition

20 mM Tris-HCl
22 mM KCl
22 mM NH4Cl
1.8 mM MgCl2
5% Glycerol
0.06% IGEPAL® CA-630
0.05% Tween® 20
0.2 mM dNTPs
1X Xylene Cyanol
1X Tartrazine
25 units/ml OneTaq Hot Start DNA Polymerase
pH 8.9@25°C

Heat Inactivation

No

Unit Assay Conditions

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

Notes

  1. Product specifications for individual components in the OneTaq Hot Start Quick-Load 2X Master Mix with Standard Buffer are available separately.
  2. OneTaq Hot Start Quick-Load 2X Master Mix with Standard Buffer is stable for fifteen freeze-thaw cycles when stored at -20°C
  3. OneTaq Hot Start Quick-Load 2X Master Mix with Standard Buffer is also stable for one month at 4°C, so for frequent use, an aliquot may be kept at 4°C.

References

  1. Barnes, W.M. (1994). Proc. Natl. Acad. Sci. USA. 91, 2216-2220.
  2. Saiki, R.K. et al. (1985). Science. 230, 1350-1354.
  3. Powell, L.M. et al. (1987). Cell. 50, 831-840.
  4. Sun, Y., Hegamyer, G. and Colburn, N. (1993). Biotechniques. 15, 372-374.
  5. Sarkar, G., Kapelner, S. and Sommer, S.S. (1990). Nucleic Acids Res.. 18, 7465.

FAQs

  1. How should I set up a PCR using the OneTaq® Hot Start Master Mixes?
  2. Can I use my regular Taq-based cycling conditions for OneTaq® Hot Start DNA Polymerase based products?
  3. How do I activate OneTaq® Hot Start Polymerase?
  4. What are the stability and storage requirements of the Quick-Load® OneTaq® Master Mixes?
  5. What type of DNA ends result from a primer extension reaction or a PCR using OneTaq® DNA Polymerase?
  6. How long a product can be made by OneTaq® DNA Polymerase?
  7. Can OneTaq® DNA Polymerase be used with uracil-containing primers or bisulfite-treated DNA?
  8. What is the fidelity of OneTaq® DNA Polymerase?
  9. Can OneTaq® DNA Polymerase be used in colony PCR?
  10. Where can I find help troubleshooting my PCR?
  11. How should I determine an appropriate annealing temperature for my reaction?
  12. How is OneTaq DNA Polymerase different from LongAmp™ Taq DNA Polymerase?

Tech Tips

A master mix with a gel loading dye is the ultimate time-saver! It allows for easy reaction set up and with the gel loading dye already in the mix, the results of your PCR are available even faster.

Protocols

  1. Protocol for OneTaq® Hot Start Quick-Load® 2X Master Mix with Standard Buffer (M0488)

Datacards

The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.

Selection Charts

Usage Guidelines & Tips

Troubleshooting Guides

Interactive Tools

Application Notes

Quality Control

Quality Control Assays

The following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page. Further information regarding NEB product quality can be found here.
  • PCR Amplification (Master Mix):
    The polymerase master mix is tested in a polymerase chain reaction (PCR) using a control template and specific primers, resulting in the expected product.

Certificate of Analysis

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

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]

Safety Data Sheet

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

Datacards

The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.