Extraction of total RNA from fresh/frozen tissue (FT)
This protocols covers the extraction of total RNA from fresh/frozen tissue. The extraction method (steps 2–21) is taken from the method supplied with TRIzol reagent (Invitrogen, Paisley, UK).
Recover tumor tissue at the time of surgery, trim into 1-cm3 fragments and immerse immediately in TRIzol reagent prior to freezing at -80°C.
Thaw and weigh tissue prior to RNA extraction, working quickly.
Use a tissue power homogenizer (or a mortar and pestle to homogenize tissue by hand).
All centrifugation steps are carried out at 4°C....
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