SeqBench

NsiI

ATGCAT3′ overhang (4)

NsiI is a 6 bp-recognition restriction enzyme. It runs best in rNEBuffer 3.1 at 37°C.

NEB buffer activity

Buffer% Activity
rNEBuffer 1.110%
rNEBuffer 2.175%
rNEBuffer 3.1best100%
rCutSmart25%
Incubation
37°C
Heat inactivation
80°C, 20 min
Star activity
None
Methylation blocked
None

ATGCAT.

Find NsiI sites in your own sequence

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

Frequently asked questions

What does NsiI recognize?

NsiI recognizes the 6 bp sequence ATGCAT (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 NsiI?

rNEBuffer 3.1 gives the highest activity (incubate at 37°C).

Does NsiI have star activity?

No documented star activity under standard reaction conditions.

Is NsiI blocked by DNA methylation?

Not blocked by Dam, Dcm, or CpG methylation.

Can I heat-inactivate NsiI?

Yes — 80°C for 20 minutes.