SeqBench

MluI

ACGCGT5′ overhang (4)

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

NEB buffer activity

Buffer% Activity
rNEBuffer 1.125%
rNEBuffer 2.175%
rNEBuffer 3.1best100%
rCutSmart50%
Incubation
37°C
Heat inactivation
65°C, 20 min
Star activity
None
Methylation blocked
CpG

ACGCGT contains CG; CpG methylation blocks.

Find MluI sites in your own sequence

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

Frequently asked questions

What does MluI recognize?

MluI recognizes the 6 bp sequence ACGCGT (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 MluI?

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

Does MluI have star activity?

No documented star activity under standard reaction conditions.

Is MluI 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 MluI?

Yes — 65°C for 20 minutes.