PCR on mouse spleen tissue

DEB THE PLEB FERGUSOND at IRAD.AFRC.AC.UK
Tue Jul 13 15:36:00 EST 1993


As part of my final year project at university I was regularly extracting
RNA from mouse spleen cells to use in PCR. The method I found the most 
succssesful was that of Chomczynski & Sacchi (1987) Single-step method of RNA
isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.
Analytical Biochemistry, volume 162 pg 156 onwards.
I simply cut out the spleen; stuck it in a glass-Teflon homogenizer with 1ml
of 'solution D' and followed the protocol.
I followed this up with a reverse transcription step and PCR for beta-actin
and gamma-IFN - worked a treat every time.
Hope this is of some use.

Debbie Ferguson
AFRC.Compton
 



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