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.
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* 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)''':
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==References==
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==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 />
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==References==
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<biblio>
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#Morrow-PNAS-1972 pmid=4343967
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// Discusses cleavage of DNA at a unique location by EcoRI
 +
#Mulder-PNAS-1972 pmid=4343959
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// Specificity of EcoRI
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#Hedgpeth-Proc-Natl-Acad-Sci-USA-1972 pmid=4343974
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// Identifies the overhang sequence produced by EcoRI
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#Bigger-Nat-New-Biol-1973 pmid=4578426
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// No abstract available online.
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#Mertz-Proc-Natl-Acad-Sci-USA-1972 pmid=4343968
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// Use of EcoRI to generate DNA fragments that can be ligated
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#Cohen-PNAS-1973 pmid=4594039
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// 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

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

  1. Morrow JF and Berg P. . pmid:4343967. PubMed HubMed [Morrow-PNAS-1972]
    Discusses cleavage of DNA at a unique location by EcoRI

  2. Mulder C and Delius H. . pmid:4343959. PubMed HubMed [Mulder-PNAS-1972]
    Specificity of EcoRI

  3. Hedgpeth J, Goodman HM, and Boyer HW. . pmid:4343974. PubMed HubMed [Hedgpeth-Proc-Natl-Acad-Sci-USA-1972]
    Identifies the overhang sequence produced by EcoRI

  4. Bigger CH, Murray K, and Murray NE. . pmid:4578426. PubMed HubMed [Bigger-Nat-New-Biol-1973]
    No abstract available online.

  5. Mertz JE and Davis RW. . pmid:4343968. PubMed HubMed [Mertz-Proc-Natl-Acad-Sci-USA-1972]
    Use of EcoRI to generate DNA fragments that can be ligated

  6. Cohen SN, Chang AC, Boyer HW, and Helling RB. . pmid:4594039. PubMed HubMed [Cohen-PNAS-1973]
    Use of EcoRI to generate recombinant DNA fragments

All Medline abstracts: PubMed HubMed
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