[Arabidopsis] BclI?

Michael Sullivan via arab-gen%40net.bio.net (by mlsulliv from wisc.edu)
Mon Mar 12 12:45:12 EST 2007


BclI is sensitive to dam methylation (dam methylation occurs at the  
sequence GATC). Most lab E. coli strains contain the dam methylase,  
so if your DNA has been prepared from a standard lab strain of E.  
coli, BclI will probably not cut it. Usually to use enzymes that are  
sensitive to dam methylation, people use special dam- strains of E.  
coli for preparing the plasmid DNA. DNA prepared in vitro (e.g., via  
a PCR reaction) should cut OK.

Hope this help.

Mike


On Mar 10, 2007, at 11:44 AM, savita from unt.edu wrote:

> Hi
>
> Could anyone please tell me if there is a specific reaction set up  
> for a BclI restriction digest? I have been trying to digest @ 50 C  
> but it is not working. Any  suggestion is most appreciated!
>
> Thanks
> Savita
>
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---
Michael L. Sullivan
Plant Research Molecular Geneticist
US Dairy Forage Research Center
ARS-USDA
1925 Linden Drive West
Madison, WI 53706
(608) 890-0046 (Phone)
(608) 890-0076 (FAX)



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