| Enzyme | Recognition Sequence (5′→3′) | Cut Site | Overhang | Overhang Seq | Heat Inact. | Buffer |
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How to Use the Restriction Enzyme Reference
Search: Type an enzyme name (e.g. EcoRI), recognition sequence (e.g. GAATTC), or overhang type (e.g. 5-prime) to filter instantly.
Filter buttons: Filter by overhang type — 5′ sticky ends, 3′ sticky ends, or blunt ends.
Click any row to see the full detail panel with recognition sequence diagram, compatible ends, methylation sensitivity, and protocol notes.
About Restriction Enzymes
Restriction endonucleases are bacterial enzymes that cut double-stranded DNA at or near specific recognition sequences. They are essential tools in molecular cloning, DNA mapping, and recombinant DNA technology.
5′ overhang (sticky): 5′—AATTC—3′ ← most common, e.g. EcoRI
3′—C —5′
3′ overhang (sticky): 5′—C —3′ ← e.g. KpnI, SacI
3′—CATGG—5′
Blunt ends: 5′—GG|CC—3′ ← e.g. SmaI, EcoRV
3′—CC|GG—5′
// ↑ = cut position on top strand
// Compatible (cohesive) ends ligate efficiently
Choosing the Right Enzyme
When planning a cloning experiment, choose enzymes that produce compatible ends for ligation. Check that the enzyme does not cut within your insert or vector sequence. Also verify that the enzyme is not blocked by Dam, Dcm, or CpG methylation if your DNA is from a methylation-positive strain.
Heat Inactivation
Many restriction enzymes can be inactivated by heating to 65°C or 80°C for 20 minutes, eliminating the need for column purification before ligation. Enzymes marked "No" or "65°C" in this table require gel or column cleanup before ligation.