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| Download: | | MSDS PDF | |
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Description:
Therminator™ γ (gamma) DNA Polymerase is a 9°Nm™ DNA polymerase variant with an enhanced ability to incorporate gamma-phosphate labeled dNTPs.
Source:
Therminator γ DNA Polymerase is purified from a strain of E. coli that carries the 9°N (D141A / E143A / W355A / L408W / R460A / Q461S / K464E / D480V / R484W / A485L) DNA Polymerase gene, a genetically engineered form of the native DNA polymerase from Thermococcus species 9°N-7. Developed by LI-COR® Biosciences and derived from Therminator.
Applications:
- Incorporation of modified nucleotides into DNA
- DNA sequencing using gamma-phosphate modified nucleotides
Therminator γ Gamma Reaction Buffer Pack (10X)
Enzyme Properties
Heat Inactivation:
No
Molecular Weight:
Theoretical: 89,580 daltons
Specific Activity:
8,500 units/mg
Reaction & Storage Conditions
Reaction Conditions:
1X Therminator γ Gamma Reaction Buffer Pack
1X Therminator γ Gamma Reaction Buffer Pack:
20 mM Tris-HCl
50 mM KCl
5 mM MgSO4
0.02 % IGEPAL
pH 9.2 @ 25°C
Unit Definition:
One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTPs into acid insoluble material in 30 minutes at 75°C.
Unit Assay Conditions: 1X Therminator γ Reaction Buffer, 200 µM dNTPs including [3H]-dTTP and 15 nM primed M13mp18.
Enzyme properties:
3´ → 5´ Exonuclease: No
5´ → 3´ Exonuclease: No
Strand Displacement: Yes
Concentration:
2,000 units/ml
Storage Conditions:
10 mM Tris-HCl
300 mM KCl
1 mM Dithiothreitol
0.1 mM EDTA
50% Glycerol
pH 7.4 @ 25°C
Storage Temperature:
-20°C
Notes
General notes:
- Reaction Conditions: 1X Therminator γ Reaction Buffer, DNA template, primer, 200 µM dNTPs and 0.5-2 units of Therminator γ DNA Polymerase in a total reaction volume of 100 µl.
Quality Control for Current Lot
Quality control values for a specific lot can be found on the datacard which accompanies each product.
Exonuclease Activity:
Incubation of a 50 μl reaction containing 20 units of Therminator™ γ DNA Polymerase with 1 μg of a mixture of single and double-stranded [3H] E. coli DNA (105 cpm/μg) for 4 hours at 37ºC released < 0.1% of the total radioactivity.
Endonuclease Activity:
Incubation of a 50 μl reaction containing 20 units of Therminator™ γ DNA Polymerase with 1 μg of ΦX174 DNA for 4 hours at 37ºC resulted in < 10% conversion to RFII as determined by agarose gel electrophoresis.
Reagents Sold Separately
Therminator γ Gamma Reaction Buffer Pack
Companion Products
Deoxynucleotide Solution Mix
Deoxynucleotide Solution Set
Diluent E
Therminator γ Gamma Reaction Buffer Pack
Legal
Patents:
New England Biolabs, Inc.: U.S. Patent No. 5,352,778
New England Biolabs, Inc.: U.S. Patent No. 5,756,334
Licensed under: PCT Publication No. W02005/037064