PstI
CTGCA▾G3′ overhang (4)
PstI is a 6 bp-recognition restriction enzyme. It runs best in rNEBuffer 3.1 at 37°C, and is prone to star activity under non-standard conditions.
NEB buffer activity
| Buffer | % Activity |
|---|---|
| rNEBuffer 1.1 | 75% |
| rNEBuffer 2.1 | 75% |
| rNEBuffer 3.1best | 100% |
| rCutSmart | 50% |
- Incubation
- 37°C
- Heat inactivation
- 80°C, 20 min
- Star activity
- High propensity
- Methylation blocked
- None
Find PstI sites in your own sequence
Paste a sequence below and this scans it for every PstI recognition site — the same engine behind the full Restriction Sites tool, scoped to just this one enzyme.
Frequently asked questions
What does PstI recognize?
PstI recognizes the 6 bp sequence CTGCAG (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 PstI?
rNEBuffer 3.1 gives the highest activity (incubate at 37°C).
Does PstI have star activity?
Yes, and it has a notably high propensity for it — under non-standard conditions (excess enzyme/glycerol, long incubation, low ionic strength) it can cleave at sites resembling but not matching CTGCAG.
Is PstI blocked by DNA methylation?
Not blocked by Dam, Dcm, or CpG methylation.
Can I heat-inactivate PstI?
Yes — 80°C for 20 minutes.