Kirsty Townsend (ktownsen at eosin.path.uwa.edu.au) writes:
>> I'm hoping someone out there can help me, because I'm having huge problems
> with restriction digests of particular strains of Pasteurella multocida.
[snip]
> I get at the end of it all, will cut with Eco RI but not with Pst I. I
> have tried using increased enzyme concentrations, different buffer etc
> etc but to no avail. Can somebody help me??? Other strains cut perfectly
> well with both enzymes, so it has to be the effect of the LPS.
>
Does it have to be the LPS? Is it possible this particular
strain carries the PstI or equivalent restriction-modification
system, so all the PstI sites are modified and won't cut for
that reason? Just a thought.
----------------------------------------------------------------------
Walter Ogston ogston at hobbes.kzoo.edu
Department of Biology Phone: (616)337-7010
Kalamazoo College Fax: (616)337-7251
Kalamazoo, MI 49006-3295