Difference between revisions of "EcoRI"

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* 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 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.
  
==References==
+
==External links==
 
[http://www.neb.com/nebecomm/products/productR0101.asp EcoRI from NEB]<br />
 
[http://www.neb.com/nebecomm/products/productR0101.asp EcoRI from NEB]<br />
 
[http://www.promega.com/catalog/catalogproducts.asp?catalog_name=Promega_Products&category_name=EcoR+I&cookie%5Ftest=1 EcoRI from Promega]<br />
 
[http://www.promega.com/catalog/catalogproducts.asp?catalog_name=Promega_Products&category_name=EcoR+I&cookie%5Ftest=1 EcoRI from Promega]<br />
 +
 +
==References==
 +
<biblio>
 +
#Hedgpeth-Proc-Natl-Acad-Sci-USA-1972 pmid=4343974
 +
// Identifies the overhang sequence produced by EcoRI
 +
#Bigger-Nat-New-Biol-1973 pmid=4578426
 +
// No abstract available online.
 +
#Mertz-Proc-Natl-Acad-Sci-USA-1972 pmid=4343968
 +
// Use of EcoRI to generate recombinant DNA fragments
 +
</biblio>
  
 
[[Category:Material]] [[Category:Enzyme]] [[Category:Endonuclease]]
 
[[Category:Material]] [[Category:Enzyme]] [[Category:Endonuclease]]

Revision as of 12:43, 20 December 2007

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.

External links

EcoRI from NEB
EcoRI from Promega

References

  1. 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. PubMed ID:4343974 | HubMed [Hedgpeth-Proc-Natl-Acad-Sci-USA-1972]
    Identifies the overhang sequence produced by EcoRI
  2. Bigger CH, Murray K, and Murray NE. Recognition sequence of a restriction enzyme. Nat New Biol. 1973 Jul 4;244(131):7-10. PubMed ID:4578426 | HubMed [Bigger-Nat-New-Biol-1973]
    No abstract available online.
  3. 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. PubMed ID:4343968 | HubMed [Mertz-Proc-Natl-Acad-Sci-USA-1972]
    Use of EcoRI to generate recombinant DNA fragments
All Medline abstracts: PubMed | HubMed