EcoRI

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==Notes==
==Notes==
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*Improving the efficiency of [[EcoRI/SpeI Double Digest]].
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* 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.  --[[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.
==References==
==References==
[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 />

Revision as of 16:41, 12 September 2006

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.

References

EcoRI from NEB
EcoRI from Promega

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