cloning

Els Van Geldre Els.VanGeldre at rug.ac.be
Tue Apr 29 03:49:46 EST 1997


Hi,

I'm trying to clone a PCR product in M13mp18 RF. After cutting the 
vector with Sma I and and blunting the 5' side of my PCR product I obtain 
few bleu plaques (I treated the vector in order to minimize selfligation) 
and much more white plaques. However, when I try to isolate ssDNA and 
sequence the insert with a universal primer on the M13 vector I 
notice that there is absolutely no insert present. I repeated the 
experiment several times, what goes wrong ?
Maybe, the problem is correlated with the fact I purify vector and PCR 
product with the Geneclean kit from Bio 101 ?
Thank you for answering,
Els




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