I want to sort cells using FACS, followed by fixation of sorted populations using formaldehyde or ethanol. I would like to extract total RNA from these sorted and fixed cells later. Has anyone done this and if yes can you share a protocol with me?
I am planning to use liver cell lines for this experiment and I am not bothered about any cell surface marker here. I would be happy if your comment is backed up by a publication or an authentic source.Thanks in advance!
Best regards,
Ankur
It depends what you want to do with the RNA. So actually the general answer is yes, you can isolate RNA from fixed cells/tissue. But it may not be useable for your experiment.
In general: Use high quality, fresh, buffered, and not stabilized PFA (we actually order now our ready-to-use PFA solution in vaccuum sealed glass bottles to ensure the quality), add additional RNAse inhibitor, use short fixation times and isolate the RNA as soon as possible.
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