Bst DNA Polymerase V2 is derived from Bacillus stearothermophilus DNA Polymerase I. It exhibits robust 5′→3′ DNA polymerase activity and strong strand displacement activity, but lacks 5′→3′ exonuclease activity. This enzyme is particularly well-suited for strand-displacement isothermal amplification methods such as LAMP (Loop-mediated Isothermal Amplification) and rapid sequencing applications.

The amount of enzyme required to incorporate 25 nmol of dNTP into acid-insoluble substances within 30 min at 65°C is defined as 1 unit of activity.

10× HH Bst V2 Buffer: 2.5 μL
MgSO4 (100 mM): 1.5 μL
dNTPs (10 mM each): 3.5 μL
SYTOTM 16 Green (25×): 1.0 μL*
Primer mix**: 6 μL
Bst DNA polymerase V2 (8 U/μL): 1 μL
DNA template: X μL
ddH2O: Up to 25 μL
*Other dyes may be substituted
**Mix includes 20 μM FIP, 20 μM BIP, 2.5 μM F3, 2.5 μM B3, 5 μM LF, 5 μM LB

Incubate in 1× HH Bst V2 Buffer at 60-65°C for 30-60 minutes.
Optional heat inactivation: 85°C for 5 minutes.

Protein Purity: ≥99% by SDS-PAGE (Coomassie Blue)
Endonuclease Activity: 8 U with 1 μg λDNA, 50 μL, 16 hrs, 37°C — no degradation
Exonuclease Activity: 8 U with 1 μg λ-Hind III digest DNA — no degradation
Nickase Activity: 8 U with 1 μg pBR322 DNA — no degradation
RNase Activity: 8 U with 1.6 μg MS2 RNA, 16 hrs at 37°C — no degradation
E. coli DNA: ≤1 copy/120 U (TaqMan qPCR, 16S rRNA primers)

Ship with ice packs. Store at -20°C. Avoid repeated freeze-thaw cycles.