4.3. Acid phenol RNA extraction from adult bees

This below recipes use an acid-phenol phase separation for isolating RNA from DNA and other tissue components. The TRIzol® (Invitrogen™) protocol is the most commonly used, and widely available, method of acid-phenol extraction of RNAs. However, using a generic lysis and acid-phenol buffer (e.g., section 4.3.2) provides a cost effective alternative than TRIzol®, and allows a great reduction in the use of the caustic chemical phenol for pooled samples. We use honey bee abdomens because they provide representation of the microbes and immune components of the honey bee, while avoiding pigments in the eye which can inhibit downstream enzymatic reactions.  The procedure is also appropriate for larvae, whole adult bees and pupal RNA extractions, if volumes are scaled upward, i.e. doubled, to reflect the volume of the sample, for the latter two.

4.3.3. RNA lysis/stabilization buffer