PCR queries

David Fogarty onxfoxxd at lgdx02.lg.ehu.es
Tue Apr 25 10:18:38 EST 1995


Hello out there,
I'm new to discussion groups and relatively new to the whole field of PCR 
so pardon the humility of my questions, if so they appear to you, kind 
reader.
1) When I run gels of my PCR products, I occasionally find a long 
"streak" in various columns rather than a neat discrete band against a 
low background.  Anyone know the reason for this?
2) I am amplifying expressed intronless genes (RT-PCR) and thus need to 
perform a DNAse treatment of my RNA from cell culture.  Any protocol 
suggestions?
3) I believe a good PCR control is to perform a "Mock RT" ie an RT 
without the enzyme.  I have done this and on occassion found a number of 
bands, although not of the predicted size.  Why is this?  Can the Taq 
polymerase act upon an RNA template in the absence of DNA?

Looking forward to hearing some suggestions,
Regards to all,
David (Department of Neuroscience)




More information about the Methods mailing list