RNAseq of Drosophila Sweet and Bitter Gustatory Second Order Neurons
RNAseq of Drosophila Sweet and Bitter Gustatory Second Order Neurons
Samples were collected after dissection in Dulbecco's Phosphate Buffer Saline, then digested by an enzymatic mixture with papaine and collagenase, and finally sorted by FACS into RNA extraction buffer. Samples were dissected in cold Calcium- and Magnesium-free 1X Dulbecco's phosphate buffered saline (DPBS), and transferred into LowBind tubes containing DPBS 0'1% bovine serum albumim (BSA). Then, samples were digested before procedure to FACS. Flies were growth in standard Iberin fly food in a 12 h light:12 h dark cycle at 25ºC until use. RNA was extracted by using the PicoArcturus (ThermoFisher, ref. 12204-01) according to manufacturer standard protocol. This procedure includes a serial centrifugation method which include washing buffer and 70% ethanol, to finally elude samples in the elution buffer provided. Libraries were prepared by the CRG (Genomic Research Centre, Barcelona, Spain) using an ultra-low RNA input protocol.