SeqBench

XbaI

TCTAGA5′ overhang (4)

XbaI is a 6 bp-recognition restriction enzyme. It runs best in rCutSmart at 37°C, and is prone to star activity under non-standard conditions.

NEB buffer activity

Buffer% Activity
rNEBuffer 1.10%
rNEBuffer 2.1100%
rNEBuffer 3.175%
rCutSmartbest100%
Incubation
37°C
Heat inactivation
65°C, 20 min
Star activity
Yes
Methylation blocked
Dam

TCTAGA. Blocked by overlapping Dam methylation (e.g., ...GATCTAGATC...). Classic Dam-sensitive enzyme.

Find XbaI sites in your own sequence

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

Frequently asked questions

What does XbaI recognize?

XbaI recognizes the 6 bp sequence TCTAGA (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 XbaI?

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

Does XbaI have star activity?

Yes — under non-standard conditions (excess enzyme/glycerol, long incubation, low ionic strength) it can cleave at sites resembling but not matching TCTAGA.

Is XbaI 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 XbaI?

Yes — 65°C for 20 minutes.