What is the effect of fixatives on RNA quality?

29 July 2014 1 521 Report

I want to Isolate RNA for sequencing. However, I first want to fix my cells. I was thinking of using 1% paraformaldehyde, is this possible?

Does this affect the quality of the RNA?

If so, which fixative is the least harmful?

Sabyasachi Biswas

Paraformaldehyde will crosslink proteins and nucleic acids and will reduce RNA yield. If you have to fix the cells try 70% ethanol which is a precipitative fixative, or acetone. You can also try PAXGene from Qiagen (tissue) or BD (bone marrow); however be aware that the PAXGene blood RNA kit will lyse your cells. Possibly the PAXGene tissue kit (Qiagen) will be a better choice.

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