DESCRIPTION
BioThermBio™ is a novel thermostable DNA polymerase that dramatically improves incorporation of Biotin- and Digoxigenin-dUTP as compared to Taq DNA polymerase.

CONCENTRATION
5 units/µl

UNIT DEFINITION
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.

STORAGE BUFFER
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)

STORAGE TEMPERATURE
Store BioThermBio™ DNA polymerase below 0ºC, preferably at -20ºC, in a constant temperature freezer.

10X REACTION BUFER
160 mM (NH₄)₂SO₄, 670 mM Tris-HCl pH 8.8 (at 25ºC), 15 mM MgCl₂, 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

ASSOCIATED ACTIVITIES
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.

Biotine-11-dUTP labelling of PCR DNA fragment (650 bp) by thermofilic DNA polymerases. Lane 1. Amplification by Taq polymerase, dNTPs 200 mM each, no Biotine-11-dUTP. Lane 2. Amplification by Taq polymerase, dNTPs 200 mM each, dTTP 130 mM, Biotine-11-dUTP 70 mM. Lane 3. Amplification by Taq polymerase, dNTPs 200 mM each, dTTP 130 mM, Biotine-11-dUTP 70 mM. Analysis with streptavidin/alcalic phsphatase conjugates shows that specific incorporation of Biotin-11-dUDP was 20 time higher withBioThermBio™ Taq DNA Polymerase.