BclI methylation sensitivity - yes or no or what?
mlsulliv at wisc.edu
Tue Mar 14 15:11:08 EST 2006
I'm not sure about your BclI problem, but I wanted to point out that
SpeI is not an isoschizomer of BclI.
Did you run out the BclI digest next to uncut plasmid?
On Mar 14, 2006, at 11:11 AM, Wolfgang Schechinger wrote:
> Dear Experts,
> BclI (SpeI) is reported to be dam methylation sensitive (see eg NEB
> Fermentas catalogues)
> just was cutting pLNCX into 2 fragments with BclI (Bcl one). As the
> plasmid was accidentally prepared in DH5alpha, actually BclI shouldn't
> have cut at all. On the gel, the digest seemed to be complete. The
> reaction was set up as usual (5 Units enzyme, 1hrs 50degC, Rxn vol 50
> What went right? Your comments are appreciated!
> Wolfgang Schechinger, PhD
> University Hospital "Bergmannheil"
> Endocrine Research
> Bochum, Germany
> PS Please reply to the NG, this email address ends in autotrash
> Methods mailing list
> Methods at net.bio.net
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)
More information about the Methods