SeqBench

MboI

GATC5′ overhang (4)

MboI 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 3.1100%
rCutSmartbest100%
Incubation
37°C
Heat inactivation
65°C, 20 min
Star activity
None
Methylation blocked
Dam

GATC. Blocked by Dam methylation (GA(me)TC) — do NOT use on Dam+ E. coli DNA. Isoschizomer DpnII also Dam-blocked; DpnI requires Dam methylation.

Find MboI sites in your own sequence

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

Frequently asked questions

What does MboI recognize?

MboI recognizes the 4 bp sequence GATC (5′→3′), cutting the top strand between position 0 and 1 within the site to leave a 5′ overhang of 4 bases.

What buffer should I use for MboI?

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

Does MboI have star activity?

No documented star activity under standard reaction conditions.

Is MboI blocked by DNA methylation?

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

Can I heat-inactivate MboI?

Yes — 65°C for 20 minutes.