EcoRI: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
Trunkflare (talk | contribs) m (→References) |
(→Notes) |
||
| Line 7: | Line 7: | ||
==Notes== | ==Notes== | ||
*Improving the efficiency of [[EcoRI/SpeI Double Digest]]. | * 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 14:41, 12 September 2006
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.
