RT PCR of Mixed Lymphocyte Cultures

[Douglas Hale]

I seem to be having a great deal of difficulty obtaining good quality RNA
from cells obtained from one way mixed lymphocyte cultures.  The 260:280
is invariably around 1.6 regardless of the amount of trizol used for the
extraction.  I am adding the trizol directly to the well (5 million
responders, 5 million irradiated stimulators) after removing most of the
media.  The specimen is then frozen at -70°C until the extraction is
carried out.  Has anyone had a similar experience?  If so, what was the
fix?

Thank you in advance.

Sincerely,

Douglas Hale