EcoRI
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* '''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. --[[User:Bcanton|BC]] 13:39, 2 Jun 2005 (EDT) | * '''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. --[[User:Bcanton|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 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. *'''[[User:Karmella Haynes|Karmella Haynes]] 14:07, 19 January 2012 (EST)''': | ||
| - | == | + | ==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> | ||
| + | #Morrow-PNAS-1972 pmid=4343967 | ||
| + | // Discusses cleavage of DNA at a unique location by EcoRI | ||
| + | #Mulder-PNAS-1972 pmid=4343959 | ||
| + | // Specificity of EcoRI | ||
| + | #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 DNA fragments that can be ligated | ||
| + | #Cohen-PNAS-1973 pmid=4594039 | ||
| + | // Use of EcoRI to generate recombinant DNA fragments | ||
| + | </biblio> | ||
[[Category:Material]] [[Category:Enzyme]] [[Category:Endonuclease]] | [[Category:Material]] [[Category:Enzyme]] [[Category:Endonuclease]] | ||
Current revision
Contents |
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.
- 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
- Morrow JF and Berg P. . pmid:4343967.
Discusses cleavage of DNA at a unique location by EcoRI - Mulder C and Delius H. . pmid:4343959.
Specificity of EcoRI - Hedgpeth J, Goodman HM, and Boyer HW. . pmid:4343974.
Identifies the overhang sequence produced by EcoRI - Bigger CH, Murray K, and Murray NE. . pmid:4578426.
No abstract available online. - Mertz JE and Davis RW. . pmid:4343968.
Use of EcoRI to generate DNA fragments that can be ligated - Cohen SN, Chang AC, Boyer HW, and Helling RB. . pmid:4594039.
Use of EcoRI to generate recombinant DNA fragments
