help with lambda phage maxi preps!
hroychow at NMSU.EDU
Fri May 19 12:14:28 EST 1995
Liquid lysates are much more difficult and time consuming to prepare. The
temperature of lysis needs to be correct and abrupt. For this you could
try to heat up equal vol of NZCYM in a water bath to about 60 C and once
the temp is reached add this to the over night culture which should also
be held at the lysis temp in a water bath. This can then be moved to a dry
shaker maintained at the lysis temp.
Alternatively, and I would recommend this to anybody, you should
try plate lysate method. If you prepare 5 or 6 plates (big or small) you
should get enough DNA to serve you for a number of manipulations. Look up
Maniatis for the protocol. It is really far simpler and yields clean DNA.
Use AGAROSE instead of AGAR for the top layer.
Hiranya S. Roychowdhury
Plant Genetic Engineering Lab.
Box 3GL, NM State Univ.
Las Cruces, NM 88003
Phone: (505) 646-5785
hroychow at nmsu.edu
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