Best way to extract RNA from FFPE human brain tissue?

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Andrey S Bavykin

Hi Antonios,

The protocol is fine as well as the reagents. However RNA yield may be dependent on specific manipulations. Use higher concentrations of PrK and longer temperature incubation (such as 3h -56 + 37 - overnight). What is the composition of your buffer for PrK? The choice of a proper paraffin also makes sense.

Franco Grimolizzi

I suggest to use the kit PureLink ™ FFPE Total RNA Isolation Kit (Life Technologies) that specifically purify RNA from FFPE tissues without the use of chemical solvents for deparaffinization. Anyway, compare yield of fresh vs FFPE and the measure relative quantity by RT-qPCR of housekeeping gene is prone to error (I would test 3 or more housekeeping genes and select for normalisation the two with the lowest SD between the fresh and the FFPE sample).