Bst DNA Polymerase, Large Fragment is the portion of the Bacillus stearothermophilus DNA Polymerase protein that contains the 5´ → 3´ polymerase activity, but lacks 5´ →3´ exonuclease activity. Enzyme provide strong strand displacement activity. The optimum temperature is from 60-65 ⁰C. Bst becomes heat inactivated at 80 ⁰C. Bst Polymerase, Large Fragment is prepared from an E. coli strain containing a gene of the Bacillus stearothermophilus DNA Polymerase gene, lacking the 5´ → 3´ exonuclease domain.
Storage condition: -20о C in 10 mM Tris-HCl (pH 8.0 at 25оС), 10 mM KCl, 1% BSA, 0,02% Tween 20, 50% glycerol.
Unit definition: One unit is defined at the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in 30 minutes at 65°C.
Quality control: Each lot of enzyme is tested for endonuclease and non specific exonuclease activity.
Areas of use
- PCR isothermal
- RNA transcription
- Whole genome sequencing