Tsg is supplied with 10 x reaction buffer and 15 mM magnesium chloride.
Tsg is a thermostable DNA Polymerase isolated from a strain of Thermus sp. It is designed for use in primer extension reaction.
Tsg has a half life of 3 hours at 95°C, it is therefore more stable than Taq DNA polymerase.
Tsg has higher fidelity than Taq with an error frequency 10e-5 to 10e-6 during DNA synthesis.

Tsg is highly purified. No detectable contaminating endonuclease, exonuclease and nicking activity is observed.

Primer Extension Characteristics: Tsg has the template-independent terminal transferase activity which results in the addition of a single nucleotide (adenosine) at 3' end of extension product. So TA cloning vector is recommended if the extension product is needed to be cloned.

Quality Testing: For endonuclease assay, 1 µg of Lambda-HindIII DNA is incubated with 20 units of the enzyme in assay buffer at 75°C for 16 hrs. and no visible contaminating activity is observed. For exonuclease assay, 1 µg of pBR322 plasmid DNA is incubated with 10 units of the enzyme for 16 hrs at 75°C in assay buffer and no detectable activity is observed. Moreover, the purity of the enzyme is also detectable by adding 10 units of Tsg DNA Polymerase in 0.1ml buffer of a reaction mixture for making first strand cDNA at beginning and no impaired effect on the first strand cDNA is observed.

Unit Definition: One unit incorporates 10 nmol of dNTP into acid-insoluble material in 30 min at 74°C.

Concentration in Storage Buffer: 5 units / µl in 20mM Tris HCl (pH 8.0), 0.1mM EDTA, 1mM DTT, 0.1% Triton X-100 and 50% glycerol.

Magnesium Chloride: The final magnesium chloride may be variable to individual requirements. In general, 1.5mM magnesium chloride in reaction mixture is workable.

Storage: -20° C