SacI
GAGCT▾C3′ overhang (4)
SacI 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 | 50% |
| rNEBuffer 3.1 | 10% |
| rCutSmartbest | 100% |
- Incubation
- 37°C
- Heat inactivation
- 65°C, 20 min
- Star activity
- None
- Methylation blocked
- None
GAGCTC contains no CG dinucleotide -> CpG not applicable.
Find SacI sites in your own sequence
Paste a sequence below and this scans it for every SacI recognition site — the same engine behind the full Restriction Sites tool, scoped to just this one enzyme.
Frequently asked questions
What does SacI recognize?
SacI recognizes the 6 bp sequence GAGCTC (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 SacI?
rCutSmart gives the highest activity (incubate at 37°C). Chart supplies NEBuffer 1 (100%); rCutSmart also 100% -> prefer rCutSmart for double digests.
Does SacI have star activity?
No documented star activity under standard reaction conditions.
Is SacI blocked by DNA methylation?
Not blocked by Dam, Dcm, or CpG methylation.
Can I heat-inactivate SacI?
Yes — 65°C for 20 minutes.