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
- CpG
GAGCTC has no internal CG, but NEB lists CpG as Blocked by Some Combinations of Overlapping — a flanking G makes the site's 3' C a CpG (GAGCTCG). Do not infer CpG insensitivity from the absence of a CG in the site itself.
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?
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 SacI?
Yes — 65°C for 20 minutes.