SmaI
CCC▾GGGBlunt
SmaI is a 6 bp-recognition restriction enzyme. It runs best in rCutSmart at 37°C.
NEB buffer activity
| Buffer | % Activity |
|---|---|
| rNEBuffer 1.1 | 0% |
| rNEBuffer 2.1 | 0% |
| rNEBuffer 3.1 | 0% |
| rCutSmartbest | 100% |
- Incubation
- 37°C
- Heat inactivation
- 65°C, 20 min
- Star activity
- None
- Methylation blocked
- CpG
Incubate at 37°C (NEB's current R0141 formulation — the classic 25°C figure is outdated). CCCGGG: CpG methylation blocks; NEB reports Dam and Dcm not sensitive. Only active in rCutSmart.
Find SmaI sites in your own sequence
Paste a sequence below and this scans it for every SmaI recognition site — the same engine behind the full Restriction Sites tool, scoped to just this one enzyme.
Frequently asked questions
What does SmaI recognize?
SmaI recognizes the 6 bp sequence CCCGGG (5′→3′), cutting the top strand between position 3 and 4 within the site to leave a blunt.
What buffer should I use for SmaI?
rCutSmart gives the highest activity (incubate at 37°C).
Does SmaI have star activity?
No documented star activity under standard reaction conditions.
Is SmaI 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 SmaI?
Yes — 65°C for 20 minutes.