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Thermophilic DNA Polymerases >
Therminator™ II DNA Polymerase |
 | | | | Therminator™ II DNA Polymerase | | | |
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Prices are in US dollars and valid only for US orders.
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- Enhanced incorporation of modified nucleotides, especially ribonucleotides
- Isolated from recombinant source
- Supplied with 10X Reaction Buffer
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).
Source:
An E. coli strain that carries the 9°N A485L/Y409V (exo-) DNA Polymerase gene, a genetically engineered form of the native DNA polymerase from Thermococcus species 9°N-7.
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)
Reagents Supplied:
ThermoPol Reaction Buffer (10X)
Enzyme Properties
Polymerase Properties | Thermophilic Polymerase Characteristics
3´ to 5´ Exonuclease: No
5´ to 3´ Exonuclease: No
Strand Displacement: Yes
Heat Inactivation:
No
Reaction & Storage Conditions
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
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.
Unit Assay Conditions: 1X ThermoPol Reaction Buffer, 200 µM dNTPs including [3H]-dTTP and 15 nM primed single-stranded M13mp18.
Concentration:
2,000 units/ml
Storage Conditions:
10 mM Tris-HCl
100 mM KCl
1 mM Dithiothreitol
0.1 mM EDTA
50% Glycerol
pH 7.4 @ 25°C
Storage Temperature:
-20°C
Quality Control for Current Lot
Quality control values for a specific lot can be found on the datacard which accompanies each product.
Quality Assurance Statement:
Purified free of contaminating endonucleases and exonucleases.
Exonuclease Activity:
Incubation of a 50 μl reaction containing 8 units of Therminator™ II DNA Polymerase with 1 μg of a mixture of single and double-stranded
[3H] E. coli DNA (205 cpm/μg) for 4 hours at 75ºC
released < 0.1% of the total radioactivity.
Endonuclease Activity:
Incubation of a 50 μl reaction containing 8 units of Therminator™ II DNA Polymerase with 1 μg of
ΦX174 RF I DNA for 4 hours at 75ºC resulted
in < 10% conversion to RFII as determined by agarose gel electrophoresis.
References
- Gardner, A.F. and Jack, W.E. (1999) Nucleic Acids Research, 27, 2545-2555.
- Gardner, A.F. and Jack, W.E. (2002) Nucleic Acids Research, 30, 605-613.
- Ruparel, H. et al. (2005) Biol.Proc. Natl. Acad. Sci., USA, 26, 5932-5937.
- Barnes, W.F. (1978) J. Mol. Biol., 119, 83-99.
- Sanger, F., Nicklen, S. and Coulson, A.R. (1977) Proc. Natl. Acad. Sci. USA, 74, 5463-5467.
- Trainor, G.L. (1996) U.S. Patent, #5, 558, 991.
- Haff, L.A. and Simirnov, I.P. (1997) Genome Methods, 7, 378-388.
Reagents Sold Separately
ThermoPol Reaction Buffer
Companion Products
9°Nm Therminator Diluent
9°Nm DNA Polymerase
Acyclonucleotide Set
Therminator™ DNA Polymerase
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