EcoRI

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

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1. Discusses cleavage of DNA at a unique location by EcoRI

2. Specificity of EcoRI

3. Identifies the overhang sequence produced by EcoRI

4. No abstract available online.

5. Use of EcoRI to generate DNA fragments that can be ligated

6. Cohen SN, Chang AC, Boyer HW, and Helling RB. Construction of biologically functional bacterial plasmids in vitro. Proc Natl Acad Sci U S A. 1973 Nov;70(11):3240-4. DOI:10.1073/pnas.70.11.3240 |

   Use of EcoRI to generate recombinant DNA fragments