SeqBench

ApoI

RAATTY5′ overhang (4)

ApoI is a 6 bp-recognition restriction enzyme. It runs best in rNEBuffer 3.1 at 50°C, and is prone to star activity under non-standard conditions.

NEB buffer activity

Buffer% Activity
rNEBuffer 1.110%
rNEBuffer 2.175%
rNEBuffer 3.1best100%
rCutSmart75%
Incubation
50°C
Heat inactivation
80°C, 20 min
Star activity
High propensity
Methylation blocked
None

RAATTY (degenerate). Incubate at 50°C for reduced star activity (ApoI* prone at 37°C). Chart lists 50°C incubation.

Find ApoI sites in your own sequence

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

Frequently asked questions

What does ApoI recognize?

ApoI recognizes the 6 bp sequence RAATTY (5′→3′), cutting the top strand between position 1 and 2 within the site to leave a 5′ overhang of 4 bases.

What buffer should I use for ApoI?

rNEBuffer 3.1 gives the highest activity (incubate at 50°C).

Does ApoI have star activity?

Yes, and it has a notably high propensity for it — under non-standard conditions (excess enzyme/glycerol, long incubation, low ionic strength) it can cleave at sites resembling but not matching RAATTY.

Is ApoI blocked by DNA methylation?

Not blocked by Dam, Dcm, or CpG methylation.

Can I heat-inactivate ApoI?

Yes — 80°C for 20 minutes.