NotI
GC▾GGCCGC5′ overhang (4)
NotI is a 8 bp-recognition restriction enzyme. It runs best in rNEBuffer 3.1 at 37°C.
NEB buffer activity
| Buffer | % Activity |
|---|---|
| rNEBuffer 1.1 | 0% |
| rNEBuffer 2.1 | 50% |
| rNEBuffer 3.1best | 100% |
| rCutSmart | 25% |
- Incubation
- 37°C
- Heat inactivation
- 65°C, 20 min
- Star activity
- None
- Methylation blocked
- CpG
GCGGCCGC has CpG dinucleotides; blocked by CpG (overlapping) methylation.
Find NotI sites in your own sequence
Paste a sequence below and this scans it for every NotI recognition site — the same engine behind the full Restriction Sites tool, scoped to just this one enzyme.
Frequently asked questions
What does NotI recognize?
NotI recognizes the 8 bp sequence GCGGCCGC (5′→3′), cutting the top strand between position 2 and 3 within the site to leave a 5′ overhang of 4 bases.
What buffer should I use for NotI?
rNEBuffer 3.1 gives the highest activity (incubate at 37°C).
Does NotI have star activity?
No documented star activity under standard reaction conditions.
Is NotI 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 NotI?
Yes — 65°C for 20 minutes.