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Thermophilic DNA Polymerases >
Therminator™ DNA Polymerase |
 | | | | Therminator™ DNA Polymerase | | | |
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Prices are in US dollars and valid only for US orders.
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- Isolated from a recombinant source
- Supplied with 10X Reaction Buffer
- Enhanced incorporation of modified nucleotides
- Single base incorporation of dideoxy and acyclonucleotides
Description:
Therminator™ 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).
Source:
An E. coli strain that carries the 9°N A485L (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 (3)
- DNA sequencing using dideoxy (4) or acyclo (5) chain terminators
- SNP analysis with dideoxy or acyclo chain terminators (6)
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
Molecular Weight:
Theoretical: 90,000 daltons
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 of 50 μl in 30 minutes at 75°C in 1X ThermoPol Reaction Buffer with 200 µM dNTPs including [3H]-dTTP and 15 nM primed single-stranded M13mp18.
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
FAQs
- What is the specific activity of Therminator™ DNA Polymerase?
- Can Therminator™ DNA Polymerase be used in other buffers?
- When should Therminator™ DNA Polymerase be used?
- Can Therminator™ DNA Polymerase be substituted for ThermoSequenase™*?
* ThermoSequenase™ is a trademark of Amersham Biosciences. - Can Therminator™ DNA Polymerase be used for sequencing?
- Why are some of the A's in my sequence weak when using Therminator™ DNA Polymerase?
- Does Therminator™ DNA Polymerase have 3'→5' proofreading exonuclease activity?
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™ 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™ DNA Polymerase with 1 μg of
ΦX174 RF I DNA for 4 hours at 37ºC resulted
in < 10% conversion to RFII as determined by agarose gel electrophoresis.
References
- Gardner, A.F. and Jack, W.E. (1999) Nucl. Acids Res., 27, 2545-2555.
- Gardner, A.F. and Jack, W.E. (2002) Nucl. Acids Res., 30, 605-613.
- 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, No. 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
Acyclonucleotide Set
Therminator™ II DNA Polymerase
ThermoPol DF (Detergent-free) Reaction Buffer
Legal
Patents:
New England Biolabs, Inc. and NEN LifeSciences (Perkin-Elmer): U.S. Patent Pending
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