SeqBench

XhoI

CTCGAG5′ overhang (4)

XhoI is a 6 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
CpG

CTCGAG contains CG; CpG methylation blocks.

Find XhoI sites in your own sequence

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

Frequently asked questions

What does XhoI recognize?

XhoI recognizes the 6 bp sequence CTCGAG (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 XhoI?

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

Does XhoI have star activity?

No documented star activity under standard reaction conditions.

Is XhoI 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 XhoI?

Yes — 65°C for 20 minutes.