DNA Polymerase, Tsg, Recombinant (Tsg), BioGenomics(TM) Kit

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 degrees C and is very stable. Tsg has high fidelity with an error frequency 10/10e6 (or 0.01/10e3) during DNA synthesis. Tsg is designed for use in primer extension reaction. Tsg (Tsg) can also be used for sequencing. DNA sequencing at high temperature may decrease the secondary structure of some DNA templates and permit polymerization through base-paired region. DNA sequencing with Tsg DNA Polymerase produces uniform band intensities and low background. Unit Definition: One unit incorporates 10nmole of dNTP into acid-insoluble material in 30min. at 74 degrees C. Kit Components: 1. Recombinant Tsg DNA Polymerase: 5 units/ul in 100mM KCl, 20mM Tris HCl (pH 8.0, 22 degrees C), 0.1mM EDTA, 0.5mM PMSF, 1mM DTT, 50% glycerol. 2. 10X Taq Reaction Buffer (Mg+2 free): (Included as D3947-01) Supplied as a liquid in 100mM potassium chloride, 100mM (NH4)2SO4, 200mM Tris HCl, pH 8.75 at RT, 1% Triton X-100, 1mg/ml BSA. Buffer is optimized for use with 200uM dNTPs. 3. Magnesium Sulfate (20mM MgSO4): (Included as D3947-02) Supplied as a liquid in 20mM MgSO4. The final magnesium sulfate may be varied 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 the 3' end of the extension product. TA cloning vector is recommended if the extension product is needed to be cloned. This product has not been licensed for PCR use. Performance & Quality Testing: Tsg DNA Polymerase is highly purified free of contaminating endonucleases, exonucleases and nicking activity. The purity of the D3947 is also evaluated by adding 10units of Tsg DNA Polymerase in 100ul of a reaction mixture for making first strand cDNA at the beginning and no impaired effect on the first strand is observed. Endonuclease Assay: 1ug of Lambda HindIII DNA is incubated with 20 units of the D3947 in assay buffer at 75 degrees C for 16 hrs. No visible contaminating activity is observed. Exonuclease Assay: 1ug of pBR322 plasmid DNA is incubated with 10 units of the D3947 in assay buffer at 75 degrees C for 16 hrs. No detectable exonuclease is observed. Storage and Stability: Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for 6 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.