Dirty PCR!

Jian_Fu Wang wang_j at MPIMG-Berlin-Dahlem.MPG.DE
Thu Apr 21 11:42:03 EST 1994


I have recently obtained two clones of the heavy chain variable region (Vh) of 
human immunoglobulin gene from a human hybridoma cell line through a standard 
procedure: RNA extraction, cDNA synthesis, PCR with published primers and 
cloning into a common-used vector. However, after sequencing, both vector and 
PCR primers are correct in  the two clones, somehow sequences of the inserts 
have no homology at all to any human Vh gene fragments and are also totally 
different from each other, though they differ to each other only three nucleic 
acids in size. It is too hard to understand it for us! We would like to know 
any similar experinces and suggestions for better understanding. Thank you in 
advance.

Jian_fu Wang
MPI for molecular biology
Berlin

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Jian_Fu Wang, Max-Planck Institut für molekulare Genetik
Ihnestrasse 73, D-14169 Berlin, Germany
e-mail: Wang_J at MPIMG-Berlin-Dahlem.MPG.DE



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