How do you clone large fragments in large vectors?

FRIEDMAN at BIOVAX.RUTGERS.EDU FRIEDMAN at BIOVAX.RUTGERS.EDU
Tue Aug 14 10:46:00 EST 1990


I have been trying for a while to clone several random fragments that I obtained
from D. melanogaster into a 16kb vector.  The fragments (6-12+kb) are already 
cloned into pUC19 vectors at the site (KpnI) which I want to use to clone into 
the 16 kb vector.  I have phosphotased the vector and it seems to not self 
ligate. The colonies I do get do not appear to be the product of the ligation. 
Any suggestions as to concentrations buffers other than standard (Maniatis) 
would be helpful
Rich
Friedman at MBCL.rutgers.edu
Friedman at biovax.rutgers.edu



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