Trypsinized cells after fixation - (Mar/10/2009 )
Do you know if I can trypsinized the cells after fixation in order to obtain RNAm for RTPCR?
I really appreciate your help.
Thanks in advance.
Claudia67 on Mar 11 2009, 02:08 AM said:
Why do you fixate cells before extracting (total) RNA?
I've always isolated total RNA from cells with a MN nucleospin column and then used a one- or two step RT kit with oligo dT primers for q-RT PCR.
Fixing crosslinks the DNA and RNA so that it will be very hard to extract.
It kind of depends on the fixation. I believe you can do it from (snap) frozen cells. Can you try lysing the cell directly. I also do that for non fixed isolations.