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Incubate at 37°C (NEB's current R0141 formulation — the classic 25°C figure is outdated).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.
Enzymes with ends compatible with SmaI
SmaI leaves blunt ends. Of the other 48 enzymes in the restriction enzyme table, these 10 leave an end that can be ligated to a SmaI end:
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.