NEBuffer EcoR I
- 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):
- Morrow JF and Berg P. Cleavage of Simian virus 40 DNA at a unique site by a bacterial restriction enzyme. Proc Natl Acad Sci U S A. 1972 Nov;69(11):3365-9. DOI:10.1073/pnas.69.11.3365 |
Discusses cleavage of DNA at a unique location by EcoRI
- Mulder C and Delius H. Specificity of the break produced by restricting endonuclease R 1 in Simian virus 40 DNA, as revealed by partial denaturation mapping. Proc Natl Acad Sci U S A. 1972 Nov;69(11):3215-9. DOI:10.1073/pnas.69.11.3215 |
Specificity of EcoRI
- Hedgpeth J, Goodman HM, and Boyer HW. DNA nucleotide sequence restricted by the RI endonuclease. Proc Natl Acad Sci U S A. 1972 Nov;69(11):3448-52. DOI:10.1073/pnas.69.11.3448 |
Identifies the overhang sequence produced by EcoRI
- Bigger CH, Murray K, and Murray NE. Recognition sequence of a restriction enzyme. Nat New Biol. 1973 Jul 4;244(131):7-10. DOI:10.1038/newbio244007a0 |
No abstract available online.
- Mertz JE and Davis RW. Cleavage of DNA by R 1 restriction endonuclease generates cohesive ends. Proc Natl Acad Sci U S A. 1972 Nov;69(11):3370-4. DOI:10.1073/pnas.69.11.3370 |
Use of EcoRI to generate DNA fragments that can be ligated
- 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