RBC lysis and RT-PCR
William L. Hanna, Ph.D.
whanna at nwu.edu
Sun Sep 22 21:23:16 EST 1996
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. I have the technique for ammonium chloride
lysis of RBCs, do not for the water lysis of RBCs (10 seconds in 9ml H2O,
add 10x PBS???).
First question: Has anyone found that the released hemoglobin after RBC
lysis (and washed away) is inhibitory for RT-PCR? If not, why do folks
use ficoll seperation? Is it to also remove the PMN which have more
Second question: What is that simple procedure for lysing RBCs with water?
Thanks for any suggestions,
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