cloning in M13

Kirk A Bartholomew kirk at NODULE.MCGILL.CA
Fri Aug 30 07:46:45 EST 1996


During my thesis work I cloned a 3.8 kb bluntended Nde1 fragment into 
the Sma1 site of M13mp18 without any serios problems.  I used both JM109 
and DH5alphaF'IQ as hosts.  I would not let your cultures overgrow when 
harvestiong phage (8-10 hours max) as this is supposed to be when you 
get rearrangements (I never saw any).  I followed standard protocols for 
cloning and harvesting DNA in M13 without any trouble.  If you have 
problems email me and I will dig out the specifics of the reactions.
good luck
kirk



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