phage display using Stratagene's Surfzap Kit
tjstbc at sd.cts.com
Mon Mar 18 11:44:48 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.
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