BclI methylation sensitivity - yes or no or what?

Michael Sullivan 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?

Mike Sullivan

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  
> and
> 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
> ul).
>
> 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
>
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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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