SeqBench

ApaI

GGGCCC3′ overhang (4)

ApaI is a 6 bp-recognition restriction enzyme. It runs best in rCutSmart at 25°C.

NEB buffer activity

Buffer% Activity
rNEBuffer 1.125%
rNEBuffer 2.150%
rNEBuffer 3.10%
rCutSmartbest100%
Incubation
25°C
Heat inactivation
65°C, 20 min
Star activity
None
Methylation blocked
CpG

GGGCCC. Incubate at 25°C. CpG methylation blocks (context).

Find ApaI sites in your own sequence

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

Frequently asked questions

What does ApaI recognize?

ApaI recognizes the 6 bp sequence GGGCCC (5′→3′), cutting the top strand between position 5 and 6 within the site to leave a 3′ overhang of 4 bases.

What buffer should I use for ApaI?

rCutSmart gives the highest activity (incubate at 25°C).

Does ApaI have star activity?

No documented star activity under standard reaction conditions.

Is ApaI blocked by DNA methylation?

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

Can I heat-inactivate ApaI?

Yes — 65°C for 20 minutes.