screening human genomic library

Xianghuai Lu lu at vh.path.uab.edu
Wed Nov 5 02:03:00 EST 1997


Hi, there:

Does anyone have experience using PCR related method to find a human
genomic clone? I know the cDNA sequences and design the primers based on
the cDNA sequences. But I use one microliter of 10-6 library boiling and
directly do PCR. I got nothing but primer dimers even using over 60C as
annealing temperuare. I also tried normal procedure using strategen
protocol by hot probe random labeling and hybridization method. It did
not work out. 

I appreciate your kindly suggestion and help.


Lu



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