Therminator™ II DNA Polymerase

  • This product was discontinued on 01/01/2013
cloned at neb recombinant unique buffer heat inactivation no
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Description

Therminator II DNA Polymerase is a 9°N™ DNA Polymerase variant with an enhanced ability to incorporate modified substrates such as dideoxynucleotides, ribonucleotides and acyclonucleotides (1,2).

Therminator II DNA Polymerase was derived from Therminator DNA Polymerase and differs in having one additional amino acid change. This change allows more efficient incorporation of ribonucleotides (1) and nucleotides with modified 3´ functional groups (3).

Highlights

  • Enhanced incorporation of modified nucleotides, especially ribonucleotides
  • Isolated from recombinant source
  • Supplied with 10X Reaction Buffer

Product Source

An E. coli strain that carries the 9°N (D141A / E143A / Y409V / A485L) DNA Polymerase gene, a genetically engineered form of the native DNA polymerase from Thermococcus species 9°N-7.

Reagents Supplied

The following reagents are supplied with this product:

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

Advantages and Features

Applications

  • DNA sequencing by partial ribosubstitution (4)
  • DNA sequencing using dideoxy (5) or acyclo (6) chain terminators
  • SNP analysis with dideoxy or acyclo chain terminators (7)

Properties and Usage

Unit Definition

One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in a total reaction volume 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
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

No

5' - 3' Exonuclease

No

3' - 5' Exonuclease

No

Strand Displacement

+

Unit Assay Conditions

1X ThermoPol Reaction Buffer, 200 µM dNTPs including [3H]-dTTP and 15 nM primed single-stranded M13mp18.

References

  1. Gardner, A.F. and Jack, W.E. (1999). Nucleic Acids Research. 27, 2545-2555.
  2. Gardner, A.F. and Jack, W.E. (2002). Nucleic Acids Research. 30, 605-613.
  3. Ruparel, H. et al. (2005). Proc. Natl. Acad. Sci., USA. 26, 5932-5937.
  4. Barnes, W.F. (1978). J. Mol. Biol.. 119, 83-99.
  5. Sanger, F., Nicklen, S. and Coulson, A.R. (1977). Proc. Natl. Acad. Sci., USA. 74, 5463-5467.
  6. Trainor, G.L. (1996). U.S. Patent 5,558,991.
  7. Haff, L.A. and Simirnov, I.P. (1997). Genome Methods. 7, 378-388.

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.
  • Exonuclease Activity (Radioactivity Release):
    The product is tested in a reaction containing a radiolabeled mixture of single and double-stranded DNA. After incubation for 4 hours the exonuclease activity is determined by the % release of radioactive nucleotides.

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.