NcoI
C▾CATGG5′ overhang (4)
NcoI is a 6 bp-recognition restriction enzyme. It runs best in rCutSmart at 37°C.
NEB buffer activity
| Buffer | % Activity |
|---|---|
| rNEBuffer 1.1 | 100% |
| rNEBuffer 2.1 | 100% |
| rNEBuffer 3.1 | 100% |
| rCutSmartbest | 100% |
- Incubation
- 37°C
- Heat inactivation
- 65°C, 20 min
- Star activity
- None
- Methylation blocked
- Dcm
CCATGG. Dcm (CCWGG) can overlap -> blocked in context.
Find NcoI sites in your own sequence
Paste a sequence below and this scans it for every NcoI recognition site — the same engine behind the full Restriction Sites tool, scoped to just this one enzyme.
Frequently asked questions
What does NcoI recognize?
NcoI recognizes the 6 bp sequence CCATGG (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 NcoI?
rCutSmart gives the highest activity (incubate at 37°C). Supplied NEBuffer 3, but 100% in all four -> rCutSmart convenient.
Does NcoI have star activity?
No documented star activity under standard reaction conditions.
Is NcoI blocked by DNA methylation?
Yes — blocked by Dcm methylation. Use methylation-free template (e.g. a dam⁻/dcm⁻ strain) if this matters for your digest.
Can I heat-inactivate NcoI?
Yes — 65°C for 20 minutes.