RT-PCR and RBC lysis (rather than ficoll)

William L. Hanna, Ph.D. whanna at nwu.edu
Mon Sep 23 18:42:44 EST 1996


Hello everyone,

The lab I've transfered into has been using ficoll seperation techniques
to remove RBCs from patient's whole blood, to look for tumor cells in the
nucleated cell fraction.

Questions:  Has anyone found that the released hemoglobin following RBC
lysis (with wash aways) is inhibitory for RT-PCR?  If not, why do folks
use ficoll seperation?  Is it to also remove the PMN which have more
RNase?

Thanks for any insights,

Bill



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