Dear Fellow Micro-Netters,
I'm hoping someone out there can help me, because I'm having huge problems
with restriction digests of particular strains of Pasteurella multocida.
They are known to have a high LPS content in the cell wall, but I thought
I was allowing for that during the million ether extractions. But the DNA
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. At present
I do 3 phenol extractions, 3 with chloroform:isoamyl alcohol, and 3 with
ether, precipitate with NaOAc and EtOH. Is there something else I should do?
Thanks in advance,
Kirsty Townsend
Molecular Oncology
QEII Medical Centre
Nedlands 6009 WA
AUSTRALIA
ktownsen at eosin.path.uwa.edu.au
