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