Unstable or unclonable cDNA

Dr. Y.H. Edwards yedwards at crc.ac.uk
Mon Oct 19 14:13:17 EST 1992


     Dear Netters,
I am writting my thesis at the moment and I need to find anyone who knows anythng about DNA which is unstable in cloning vectors.  I have cloned a region of 
cDNA which was only detectable in an unamplified library and was impossible
to maxiprep or subclone.  I have sequenced it directly from PCR product and now 
am in the position to examine the DNA sequence and to suggest reasons for the 
difficulty in retention of the fragment in a vector.  I have run a GCG 'dotplot'
with the sequence compared to itself, but this reveals no obvious repeat regioand I have tried a 'FoldRNA' run.  although this looks pretty I'm not sure how
much it means and also I have 5Kb to examine and this program can only cope
with 1.2Kb at a time.
If people have any advice or any references that they could give me I would be eternally grateful!!
To all those doing their Ph.D., hang on in there!!
                           Thanks in advance,
                           Una Fairbrother
                           HBGU, UCL
                           4 Stephenson Way
                             London NW1 2HE
                             yedwards. at uk.ac.crc
P.S. Thats my boss' mail file so be nice!!!




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