TA cloning RED-- HELP!

Daniel Miller miller.1532 at osu.edu
Wed Sep 4 18:00:26 EST 1996


I am trying to TA clone a PCR product into a pCR2.1 from Invitrogen. My 
ligation and transformations are perfect but when I do a plasmid prep 
and try to analyze the clones for the orientation of my PCR product, 
there will be no or very little plasmid in my ethidium/agarose gel. I 
perform an analagous RED on all PCR products prior to ligation and do 
ODs on recovered clones. So somewhere between ODs, the restriction 
enzyme digest reaction, and loading the RED samples onto a gel I am 
losing my clones. Please help.





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