SSDNA production via phagemids

P. Knygsman fred at gpu.utcc.utoronto.ca
Sun Feb 14 11:59:41 EST 1993


Dear Netters;
This is my first posting venture, please forgive any netting
blunders!
Does anyone have tricks on getting reproducible SSDNA yields in
phagemid mediated SSDNA production - other than the basics found
in Maniatis and Current Protocols?
We use DH5`F' cells, R408 helper, T7T3 phagemids, and a variety
of inserts.  We look at culture supernatants before purifying
SSDNA to see whether it is worth purifying a particular batch and
find that even with the same clone, upon culturing up different
batches, will give variable amounts of phagemid SSDNA (and
sometimes will have no phagemid SSDNA yielded).
Any tips would be greatly appreciated.  Please send replies by
Email to fred at utcc.utoronto.ca  Thanks for your time :)          



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