KpnI
GGTAC▾C3′ overhang (4)
KpnI is a 6 bp-recognition restriction enzyme. It runs best in rNEBuffer 1.1 at 37°C, and is prone to star activity under non-standard conditions.
NEB buffer activity
| Buffer | % Activity |
|---|---|
| rNEBuffer 1.1best | 100% |
| rNEBuffer 2.1 | 75% |
| rNEBuffer 3.1 | 0% |
| rCutSmart | 50% |
- Incubation
- 37°C
- Heat inactivation
- Not inactivable
- Star activity
- Yes
- Methylation blocked
- CpG
GGTACC. Not heat-inactivable. CpG methylation blocks (context).
Find KpnI sites in your own sequence
Paste a sequence below and this scans it for every KpnI recognition site — the same engine behind the full Restriction Sites tool, scoped to just this one enzyme.
Frequently asked questions
What does KpnI recognize?
KpnI recognizes the 6 bp sequence GGTACC (5′→3′), cutting the top strand between position 5 and 6 within the site to leave a 3′ overhang of 4 bases.
What buffer should I use for KpnI?
rNEBuffer 1.1 gives the highest activity (incubate at 37°C).
Does KpnI 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 GGTACC.
Is KpnI 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 KpnI?
No — KpnI is not heat-inactivable. Use a column cleanup or phenol-chloroform extraction to stop the reaction instead.