rt-pcr problems

Jose M. Bautista jbautist at hgmp.mrc.ac.uk
Tue Jan 2 11:18:22 EST 1996

On 1 Jan 1996, William Sun wrote:

> Hello all,
> I have been trying to amplify messages using RT-PCR, but I don't get any
> bands after the PCR.  I failed to see bands even with primers from 
> Clontech designed to amplify G3PDH.  The protocol that came with the
> primers recommended that I use oligo-dT or random primers during the RT
> step, but I just used the 3' PCR primer.  Has anyone out there have
> success with RT-PCR without using oligo-dT or random priming?  I don't 
> understand why random or oligo-dT priming would give better results.
> -William

How long did you incubate the RT reaction? If less than 1 hour here you
have your trouble: not enough lenght extension of the 1st strand to
amplify by PCR with the second primer.

Hope it help. If not, please, give more details.


More information about the Methods mailing list