BioTherm™ DNA polymerase is a thermostable Taq DNA polymerase purified from the Thermus aquaticus strain in accordance with the procedures developed by Kaledin (1,2,3) for the isolation of thermostable enzymes processing DNA polymerase activity from thermophilic bacteria. Amplification of DNA fragments (100 bp to 10kb) can be achieved with this enzyme. The enzyme has both 5'-3' polymerase- and 5'-3' exonuclease activities.
BioTherm™ can add a single template-directed deoxyadenosin (A) residue to the 3’ end of duplex PCR products.
One unit is defined as the amount of enzyme that incorporates 10 nmoles of dNTPs into acidinsoluble form in 30 minutes at 72ºC under the assay conditions (25 mM TAPS (tris-(hydroxy-methyl)-methyl-amino-pro-panesulfonic acid, sodium salt) pH 9.3 (at 25ºC); 50 mM KCl; 2 mM MgCl2; 1 mM ß-mercaptoethanol) and activated calf thymus DNA as substrate.
10 mM K-phosphate buffer pH 7.0, 100 mM NaCl, 0.5 mM EDTA, 1 mM DTT, 0.01% Tween 20, 50% glycerol(v/v)
Store BioTherm™ DNA polymerase below 0ºC, preferably at -20ºC, in a constant temperature freezer.
10X REACTION BUFER
160 mM (NH 4 ) 2 SO 4 , 670 mM Tris-HCl pH 8.8 (at 25ºC), 15 mM MgCl 2 , 0.1% Tween 20
The 10x reaction buffer (on request with or without MgCl2) is delivered free of charge.
1.5 ml 10x reaction buffer (contains 15 mM MgCl2) Cat. No. GC-002-006
1.5 ml 10x reaction buffer without MgCl2 plus 50 mM MgCl2 separately Cat. No. GC-002-007
Endonuclease- and exonuclease activities were not detectable after 2 and 1 hours incubation, respectively, of 1 µg lambda DNA and 0.22 µg of EcoRI digested lambda DNA, respectively, at 72ºC in the presence of 15 - 20 units of BioTherm™ DNA polymerase.
BioThermRed™ DNA polymerase, SupraTherm™ DNA polymerase, dNTPs, T-Vector
1 Kaledin, A.S., et al. (1980) Biokhimiya 45, 494
2 Kaledin, A.S., et al. (1981) Biokhimiya 45, 1576
3 Kaledin, A.S., et al. (1982) Biokhimiya 47, 1785