EcoRI

From OpenWetWare

Properties

Recognition site

Buffers

NEBuffer EcoR I

Notes

• Improving the efficiency of EcoRI/SpeI Double Digest.
• Reaction Volumes - I used to have very inconsistent results when digesting with EcoRI, using the 20μl reaction volume. Using the Knight Lab's 50μl mix has recently given good results. I haven't tested this enough to say it is a statistically significant result. --BC 13:39, 2 Jun 2005 (EDT)
• The custom EcoRI buffer provided by NEB contains Triton-X100. This detergent is very inhibitory in transformations, so direct ligation and cloning of DNA cut in this buffer is problematic. Remarkably small amounts of this buffer dramatically reduces transformation efficiency.

External links

EcoRI from NEB
EcoRI from Promega

References

1. Hedgpeth J, Goodman HM, and Boyer HW. . pmid:4343974. PubMed HubMed [Hedgpeth-Proc-Natl-Acad-Sci-USA-1972]

   Identifies the overhang sequence produced by EcoRI

2. Bigger CH, Murray K, and Murray NE. . pmid:4578426. PubMed HubMed [Bigger-Nat-New-Biol-1973]

   No abstract available online.

3. Mertz JE and Davis RW. . pmid:4343968. PubMed HubMed [Mertz-Proc-Natl-Acad-Sci-USA-1972]

   Use of EcoRI to generate recombinant DNA fragments