phage injection answer

SternM Lab SternM.Lab at QUICKMAIL.CIS.YALE.EDU
Mon May 15 17:55:56 EST 1995


                       Subject:                               Time:6:46 PM
  OFFICE MEMO          phage injection answer                 Date:5/15/95

A while ago, I asked how whether there were any tricks (ie concentration, prep
protocols etc.) to using phage DNA in injections.  I received one answer from
Raymond Lee:

I was trying to rescue eat-5 with some genomic phage clones Leon had fished out
from a Stratagene library (lambda FIX II vector).  DNA was prepped from liquid
lysate (the protocol is similar to yours), injected at 50ng/ul phage DNA +
50ng/ul pRAK3 DNA in TE(pH8) buffer.  Pooling data from 3 different phages, out
of 34 worms injected, I got 5 F2 lines.  One phage gave me one line and it
rescued.  I started to subclone that one into plasmids.(pRAK3 is essentially
pRF4 but with a Kan-R inserted in.)

Thanks for your answer.
Laura Selfors





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