EcoRI
G▾AATTC5′ overhang (4)
EcoRI is a 6 bp-recognition restriction enzyme. It runs best in its own unique NEB buffer at 37°C, and is prone to star activity under non-standard conditions.
NEB buffer activity
| Buffer | % Activity |
|---|---|
| rNEBuffer 1.1 | 25% |
| rNEBuffer 2.1 | 100% |
| rNEBuffer 3.1 | 50% |
| rCutSmart | 50% |
- Incubation
- 37°C
- Heat inactivation
- 65°C, 20 min
- Star activity
- High propensity
- Methylation blocked
- CpG
Classic star-activity enzyme (EcoRI*); NEB flags star activity in r2.1 or rCutSmart. GAATTC has no internal CG, but NEB reports CpG blocking for some combinations of overlapping methylation.
Find EcoRI sites in your own sequence
Paste a sequence below and this scans it for every EcoRI recognition site — the same engine behind the full Restriction Sites tool, scoped to just this one enzyme.
Frequently asked questions
What does EcoRI recognize?
EcoRI recognizes the 6 bp sequence GAATTC (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 EcoRI?
EcoRI is supplied in its own unique NEB buffer. Supplied in unique NEBuffer EcoRI (100% there). In the standard set only r2.1 is 100%; r1.1=25, r3.1=50, rCutSmart=50.
Does EcoRI 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 GAATTC.
Is EcoRI 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 EcoRI?
Yes — 65°C for 20 minutes.