Direct PCR from E. coli colonies

B VISSER * x2818 Plantkunde * pbv at rs.uovs.ac.za
Sun Jul 9 03:46:50 EST 1995


Hi Netters

Just a quick question.  I recently PCR amplified cDNA fragments directly 
from plaques by first incubating the phage particles at 70oC for 10 minutes 
to disrupt the capsid protein.  This liberated the phage DNA which could 
then be used for the amplification.  The amplified DNA was digested and then 
cloned into a shuttle plasmid vector.

I am now testing whether the digested cDNA was successfully cloned into the 
plasmid vector, but instead of minipreps, I want to amplify the inserts 
directly from E. coli colonies.  Can the same protocol used for the phage 
DNA be used directly for amplification of DNA from these colonies?  If not, 
could you suggest a publication where I could find a suitable protocol?  
Thanks in advance.

Botma Visser
pbv at rs.uovs.ac.za



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