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.
On Mar 10, 2007, at 11:44 AM, savita from unt.edu wrote:
>> 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!
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Michael L. Sullivan
Plant Research Molecular Geneticist
US Dairy Forage Research Center
1925 Linden Drive West
Madison, WI 53706
(608) 890-0046 (Phone)
(608) 890-0076 (FAX)