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Description:
Tth DNA polymerase is a thermostable 93 kDa single-subunit enzyme purified from the E. coli recombinant strain expressing Thermus thermophilus HB-8 DNA polymerase gene. Amplification of target DNA fragments from 100 bp to 3000 bp can be achieved with this enzyme. Tth DNA polymerase also exhibits reverse transcription activity in presence Mn²⁺ ions much higher than that demonstrated for Taq DNA polymerase.

Storage and dilution buffer:
10 mM K phosphate buffer, pH 7.0, 100 mM NaCl, 0.5 mM EDTA, 1 mM DTT, 50 % glycerol, 0.1 mg/ml BSA.

Unit definition:
One unit is defined as the amount of enzyme that incorporates 10 nmol of total dNTP’s into acid-insoluble form in 30 minutes at 70 ^oC under standard assay conditions.

Amplification buffer 10x, RT-PCR buffer 10x:
NH₄-buffer: 166 mM (NH₄)₂SO₄, 670 mM Tris-HCl (pH 8.8 at 25 °C), 0.1 % Tween-20.

Chelating buffer 10x (for RT-PCR):
166 mM (NH₄)₂SO₄; 670 mM Tris-HCl (pH 8.8 at 25 °C); 0.25 % Tween-20; 7.5 mM EGTA; 50 % glycerol.

Applications:
Reverse transcription: synthesizes DNA in the 5′-3′ direction from RNA template.
Primer extension: polymerizes dNTP’s in the 5′-3′ direction on single-stranded DNA template.

Quality control:
Activity, SDS-PAGE purity; endonuclease/nickase and exonuclease activities were not detectable under standard assay condition.

Concentration:
5 units/µl

Storage:
-20 °C

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