phage display using Stratagene's Surfzap Kit
lpss at unixg.ubc.ca
Mon Mar 18 18:46:47 EST 1996
: Alex, We have been using this system regularly and, on average, obtain
: primary libraries in the 10E7 range. We use either Gigapack Gold III
: extract or Epicentre extract (the latter is a lot cheaper and has
: comparable efficiencies). We habitually digest with Not/Spe overnight
: since Not1 is not the most efficient enzyme in the world. Do you have a
: long (8+ bases) overhang on the Not1 end? We also tend to ligate
: overnight at 4oC. Our background seems to vary between 10E2 and 10E3.
: Hope some of this is useful. Barry.
I did all that you've mentioned, plus gel purification after
restriction digestion using Qiagen kit. (another note: our's is
Gigapack Gold II extract, will that make any difference?)
I am wondering whether the ratio of the insert vs lambda arms is
important. That's the only thing I can think of (contributing to the
wrong ligation --> low package efficiency). Any thoughts?
University of British Columbia
achang at hivnet.ubc.ca
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