EcoRI

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Properties

Recognition site

http://www.neb.com/nebecomm/productfiles/314/images/EcoR-I-cutsite_1.gif

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.
  • The FastDigest restriction enzymes from Fermentas all work very efficiently in one universal buffer. I perform digests in a 30 μL reaction volume. *Karmella Haynes 14:07, 19 January 2012 (EST):

External links

EcoRI from NEB
EcoRI from Promega

References

<biblio>

  1. Morrow-PNAS-1972 pmid=4343967

// Discusses cleavage of DNA at a unique location by EcoRI

  1. Mulder-PNAS-1972 pmid=4343959

// Specificity of EcoRI

  1. Hedgpeth-Proc-Natl-Acad-Sci-USA-1972 pmid=4343974

// Identifies the overhang sequence produced by EcoRI

  1. Bigger-Nat-New-Biol-1973 pmid=4578426

// No abstract available online.

  1. Mertz-Proc-Natl-Acad-Sci-USA-1972 pmid=4343968

// Use of EcoRI to generate DNA fragments that can be ligated

  1. Cohen-PNAS-1973 pmid=4594039

// Use of EcoRI to generate recombinant DNA fragments </biblio>