HindIII
A▾AGCTT5′ overhang (4)
HindIII is a 6 bp-recognition restriction enzyme. It runs best in rNEBuffer 2.1 at 37°C, and is prone to star activity under non-standard conditions.
NEB buffer activity
| Buffer | % Activity |
|---|---|
| rNEBuffer 1.1 | 50% |
| rNEBuffer 2.1best | 100% |
| rNEBuffer 3.1 | 10% |
| rCutSmart | 50% |
- Incubation
- 37°C
- Heat inactivation
- 65°C, 20 min
- Star activity
- High propensity
- Methylation blocked
- None
Find HindIII sites in your own sequence
Paste a sequence below and this scans it for every HindIII recognition site — the same engine behind the full Restriction Sites tool, scoped to just this one enzyme.
Frequently asked questions
What does HindIII recognize?
HindIII recognizes the 6 bp sequence AAGCTT (5′→3′), cutting the top strand between position 1 and 2 within the site to leave a 5′ overhang of 4 bases.
What buffer should I use for HindIII?
rNEBuffer 2.1 gives the highest activity (incubate at 37°C).
Does HindIII 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 AAGCTT.
Is HindIII blocked by DNA methylation?
Not blocked by Dam, Dcm, or CpG methylation.
Can I heat-inactivate HindIII?
Yes — 65°C for 20 minutes.