Unstable or unclonable cDNA

bpn0802 at cc.utah.edu bpn0802 at cc.utah.edu
Tue Oct 20 22:39:03 EST 1992


In article <1992Oct19.191014.10871 at gserv1.dl.ac.uk>, yedwards at crc.ac.uk (Dr. Y.H. Edwards) writes:
> 
>      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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