SeqBench

MspI

CCGG5′ overhang (2)

MspI is a 4 bp-recognition restriction enzyme. It runs best in rCutSmart at 37°C.

NEB buffer activity

Buffer% Activity
rNEBuffer 1.175%
rNEBuffer 2.1100%
rNEBuffer 3.150%
rCutSmartbest100%
Incubation
37°C
Heat inactivation
Not inactivable
Star activity
None
Methylation blocked
None

CCGG. Notably CpG-METHYLATION TOLERANT (cuts C(5me)CGG) — its isoschizomer HpaII is blocked. Not heat-inactivable.

Find MspI sites in your own sequence

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

Frequently asked questions

What does MspI recognize?

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

What buffer should I use for MspI?

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

Does MspI have star activity?

No documented star activity under standard reaction conditions.

Is MspI blocked by DNA methylation?

Not blocked by Dam, Dcm, or CpG methylation.

Can I heat-inactivate MspI?

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