Tsg DNA Polymerase, 5u/ul: Tsg DNA Polymerase, 5u/ul
Product information Tsg DNA Polymerase (New Buffer!)
Tsg DNA Polymerase is a thermostable DNA Polymerase isolated from a strain of Thermus sp. Tsg has a half life of 3 hours at 95°C, it is therefore more stable than many other DNA Polymerases. Tsg has high fidelity with an error frequency 10/10^6 (or 0.01/10^3) during DNA synthesis.
Tsg is designed for use in primer extension reaction. Tsg can also be used for sequencing. DNA sequencing at high temperature may decrease the second structure of some DNA templates and permit polymerization through base-paired region. DNA sequencing with Tsg DNA Polymerase produces uniform bands intensities and low background.
Performance & Quality Testing:
Tsg DNA Polymerase is highly purified free of contaminating endonucleases, exonucleases and nicking activity.
For endonuclease assay, 1 ug of Lambda /Hind III 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 exonucleases assay, 1ug of pBR322 plasmid DNA is incubated with 10 units of enzyme for 16 hrs at 75°C in assay buffer and no detectable exonuclease is observed.
The purity of the enzyme is also evaluated by adding 10 units of Tsg DNA Polymerase in 100ul of a reaction mixture for making first strand cDNA at beginning and no impaired effect on the first strand is observed.
One unit incorporates 10nmole of dNTP into acid-insoluble material in 30 min. at 74°C.
Concentration in Storage Buffer:
5 units/ul in 100mM KCl, 20mM Tris HCl ( pH 8.0, 22°C ), 0.1mM EDTA, 0.5mM PMSF, 1mM DTT, 50% glycerol.
10 X Tsg Reaction Buffer:
100mM KCl, 100mM (NH4)2SO4, 200mM Tris HCl (pH 8.75) at 22°C, 1% Triton X-100 and 1mg/ml BSA. Buffer is optimized for use with 200uM dNTPs.
20mM MgSO4. The final magnesium sulfate may be variable according to individual requirements. In general, 2mM MgSO4 is recommended.
Primer Extension Characteristics:
Tsg has the independent terminal transferase activity which results in the addition of a single nucleotide (adenosine) at 3' end of the extension product. TA cloning vector is recommended if the extension product is needed to be cloned.