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
- None
Classic star-activity enzyme (EcoRI*); NEB flags star activity in r2.1 or rCutSmart. GAATTC has no Dam/Dcm/CpG overlap.
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?
Not blocked by Dam, Dcm, or CpG methylation.
Can I heat-inactivate EcoRI?
Yes — 65°C for 20 minutes.