urgent: strange(??) 5 race problem

Tasos Moressis pushertm at yahoo.com
Thu Jun 6 06:20:59 EST 2002


Mathias,
Thanks a lot for your suggestion! Actually I had a similar idea and I
wanted to ask whether this would be possible with cDNA.? This means,
reverse transcription (with poly-T primer), circulrization of cDNA
molecule , and pcr with primers designed for 5' and 3' Race from
central region of gene of interest.(sth like i-pcr). The problem is ,I
think, that self-ligation of ssDNA is not possible. Is there a way to
make it double stranded? (for example , repeating reverse transciption
with a poly-A primer - I read that reverse trancriptase uses cDNA with
about same efficiency as RNA...).
It would be very interesting and helpful to obtain both cDNA ends from
a single reaction or a single cDNA template...!
Thanks in advance

> > Tasos Moressis
> > PLant Molecular Biology Laboratory
> > Dept. of Agricultural Biotechnology
> > Agricultural University of Athens
> > Greece




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