HELP -DNA extraction from phage

jacqueline heard jheard at hermes.bc.edu
Wed Jul 26 12:03:06 EST 1995


Hi netsurfers and gene jocks,

	I am in need of new insights and/or protocols to isolate DNA from
bacteriophage.  
I have used a phage kit from Bio 101 and my results were far from 
exemplary.  The protocol I am currently using has allowed me to get
excellent DNA
yields but the purity is subpar, mucho degradation.  The degradation is
preventing me from subcloning.  The procedure calls for
precipitating the phage with PEG and NaCl,
chilling for one hour, and then spinning at 10,000rpms for fifteen
minutes.  After
resuspending the pellets in SM, the protocol calls for phenol/chloroforms
until the interphase is devoid of any residual phage proteins. From there
I do an ethanol 
precip. and wash.  I have minimalized the time that the DNA is in contact
with the buffered  phenol by doing repeated one mintue aggitations and
spins until the
interphase is clean.  However, to no avail, this hasn't prevented my
degradation.
If anyone out there has any advice or new strategies, I would greatly
appreciate
hearing them. I'm back. I'm back.

Respectfully yours,

John Lesher








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