Taq DNA Polymerase with ThermoPol® Buffer

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cloned at neb recombinant unique buffer heat inactivation no pcr tm-5
Catalog #SizeConcentrationPriceQtyAdd to Cart
M0267S400 units5,000 units/ml$61.00Add to Cart
M0267L2,000 units5,000 units/ml$244.00Add to Cart
M0267X4,000 units5,000 units/ml$443.00Add to Cart
M0267E20,000 units5,000 units/ml$1,200.00Add to Cart
  
Categories:
Taq DNA Polymerase Products,
Taq DNA Polymerase Products
Applications:
A-tailing,
Colony PCR,
Multiplex PCR

Description

  
Taq DNA Polymerase is a thermostable DNA polymerase that possesses a 5´→3´ polymerase activity (1,2,3) and a 5´ flap endonuclease activity (4,5).

It is supplied with 10X ThermoPol Reaction Buffer, which contains a nonionic detergent to increase enzyme stability during longer incubations.

Highlights

  • Isolated from a recombinant source
  • Supplied with 10X Reaction Buffer
  • Robust and reliable reactions
  • Tolerates a wide range of templates
  • Incorporates dUTP, dITP and fluorescently-labeled nucleotides
  • Exceptional value in terms of cost per unit

Product Source

An E. coli strain that carries the Taq DNA Polymerase gene from Thermus aquaticus YT-1

Reagents Supplied

The following reagents are supplied with this product:

Store at (°C)Concentration
ThermoPol® Reaction Buffer Pack-2010X

Advantages and Features

Applications

  • PCR
  • Primer extension
  • Colony PCR

Properties and Usage

Unit Definition

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

Reaction Conditions

1X ThermoPol® Reaction Buffer

1X ThermoPol® Reaction Buffer:
20 mM Tris-HCl
10 mM (NH4)2SO4
10 mM KCl
2 mM MgSO4
0.1% Triton® X-100
pH 8.8 @ 25°C

Storage Temperature

-20°C

Storage Conditions

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

Strand Displacement

+1

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. 

Error Rate

~ 285x10-6bases

Notes

  1. 5'→3' flap endonuclease activity destroys displaced strand.

References

  1. Chien, A., Edgar, D.B. and Trela, J.M. (1976). J. Bact.. 127, 1550-1557.
  2. Kaledin, A.S., Sliusarenko, 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.
  9. Sarkar, G., Kapelner, S. and Sommer, S.S. (1990). Nucleic Acids Res.. 18, 7465.

FAQs

  1. Can Taq DNA Polymerase be used in other buffers?
  2. Why is the product a smear when visualized on an agarose gel?
  3. Why is there no product when visualized on an agarose gel?
  4. How should I set up an amplification reaction using Taq DNA Polymerase?
  5. What type of DNA ends result from a primer extension reaction or a PCR using Taq DNA Polymerase?
  6. The product sequence doesn't completely match the expected sequence. How can this result be improved?
  7. When should Taq DNA Polymerase be used in a primer extension reaction or for PCR?
  8. Will the 5'→3' flap endonuclease activity of Taq DNA Polymerase degrade primers?
  9. Can Taq DNA Polymerase be used for nick translation?
  10. Does the presence of Ca2+ inhibit PCR?

Tech Tips

Did you know most Taq reactions amplify more efficiently and robustly when you use a 68°C extension temperature?

Protocols

  1. PCR Protocol for Taq DNA Polymerase with ThermoPol® Buffer (M0267)
  2. A-Tailing with Taq Polymerase

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

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.
  • Endonuclease Activity (Nicking):
    The product is tested in a reaction containing a supercoiled DNA substrate. After incubation for 4 hours the percent converted to the nicked form is determined by agarose gel electrophoresis.
  • 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.
  • Single Stranded DNase Activity (FAM Labeled Oligo):
    The product is tested in a reaction containing a fluorescent internal labeled single stranded oligonucleotide. The percent degradation is determined by capillary electrophoresis.

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.