RTPCR to PCR

C.R.Baker crbaker at mail.tcd.ie
Sat Dec 14 09:35:18 EST 1996


One year ago a fellow lab member amplified a cDNA from rat liver mRNA 
of approx. 670bp and kept a few of the low amplification bands at -20 
in the form of the 'cut out' agarose block following electrophoresis 
in TAE. We have just tried to extract the DNA from the gel blocks and 
PCR amplify using the same primers and programme (minus RT step) but 
without success. Buffer dNTPs MgCl2 and taq are o.k. We've used 
Geneclean, BM agarose extraction kit, and centrifugation through 
filter paper to isolate some template. Are there any reasons why the 
PCR should not work? and is there anything else to try?
Thanks for any suggestions.

Cara Baker,
Dept. of Biochemistry
TCD.



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