Homogenize 1 g adipose tissue in 5 mL of cold TRIzol solution for 30s.
Centrifuge at 2600 x g for 30 min at 2 °C.
Remove excess fat layer.
Transfer cleared homogenate to clean tube.
Incubate for 5 min. at 22 °C.
Add 0.2 ml of chloroform per ml of TRIzol reagent.
Cap tubes and shake vigorously for 15s.
Incubate for 2–3 min. at 22 °C.
Centrifuge at 2600 x g for 30 min.
Transfer clear aqueous layer to clean tube.
Add 0.57x the amount of aqueous layer in isopropanol to the tube and mix.
Load onto PureYield™ RNA Midiprep System clearing column (Promega).
Elute under vacuum.
Wash with 20 mL RNA wash solution under vacuum.
Place column in clean 50 mL tube and 1 ml of RNase-free water to top of column.
Centrifuge at 2000 x g for 3 min.
Quantify RNA. Approximate yield of 35 ug/g of subcutaneous bovine adipose tissue with 260/280 ratios of >1.8 using this method.