SeqBench

BstBI

TTCGAA5′ overhang (2)

BstBI is a 6 bp-recognition restriction enzyme. It runs best in rCutSmart at 65°C.

NEB buffer activity

Buffer% Activity
rNEBuffer 1.175%
rNEBuffer 2.150%
rNEBuffer 3.125%
rCutSmartbest100%
Incubation
65°C
Heat inactivation
Not inactivable
Star activity
None
Methylation blocked
Dam, CpG

TTCGAA. Incubate at 65°C (thermophilic). Not heat-inactivable. Dam-overlap and CpG block.

Find BstBI sites in your own sequence

Paste a sequence below and this scans it for every BstBI recognition site — the same engine behind the full Restriction Sites tool, scoped to just this one enzyme.

Frequently asked questions

What does BstBI recognize?

BstBI recognizes the 6 bp sequence TTCGAA (5′→3′), cutting the top strand between position 2 and 3 within the site to leave a 5′ overhang of 2 bases.

What buffer should I use for BstBI?

rCutSmart gives the highest activity (incubate at 65°C).

Does BstBI have star activity?

No documented star activity under standard reaction conditions.

Is BstBI blocked by DNA methylation?

Yes — blocked by Dam, CpG methylation. Use methylation-free template (e.g. a dam⁻/dcm⁻ strain) if this matters for your digest.

Can I heat-inactivate BstBI?

No — BstBI is not heat-inactivable. Use a column cleanup or phenol-chloroform extraction to stop the reaction instead.