SphI
GCATG▾C3′ overhang (4)
SphI 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 | 100% |
| rNEBuffer 3.1 | 50% |
| rCutSmartbest | 100% |
- Incubation
- 37°C
- Heat inactivation
- 65°C, 20 min
- Star activity
- None
- Methylation blocked
- None
GCATGC.
Find SphI sites in your own sequence
Paste a sequence below and this scans it for every SphI recognition site — the same engine behind the full Restriction Sites tool, scoped to just this one enzyme.
Frequently asked questions
What does SphI recognize?
SphI recognizes the 6 bp sequence GCATGC (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 SphI?
rCutSmart gives the highest activity (incubate at 37°C). Supplied NEBuffer 2; rCutSmart 100% too.
Does SphI have star activity?
No documented star activity under standard reaction conditions.
Is SphI blocked by DNA methylation?
Not blocked by Dam, Dcm, or CpG methylation.
Can I heat-inactivate SphI?
Yes — 65°C for 20 minutes.