GSM8808453: CIN brain 6; Drosophila melanogaster; RNA-Seq
1) Thaw lysis buffer (Functional Genomics Core Facility, FGCF) and confirm there are no precipitated salts. If so, vortex and spin. 2) Add 45 μL of lysis buffer into each Eppendorf (1.5 mL RNAse-free Eppendorf). Maintain the tube embebed in ice. 3) Select the larvae of interest and keep them in PBS in a well on ice.4) Dissect the tissue from the larvae in PBS and place the tissue fragment in the lysis buffer. Homogenize the tissue 5-10 times with the pipette. 5) Close the lid of the tube, perform a quick spin and incubate for 15 min at 65ºC in heat-block or a water bath. 6) After the incubation, spin down the tubes to recover condensates. 7) Save immediately the tubes at -20ºC/ -80ºC for submission to the FGCF. RNA libraries were prepared for sequencing using the kits NEBNext® Poly(A) mRNA Magnetic Isolation Module (Cat# E7490) and NEBNext® Ultra™ II RNA Library Prep Kit for Illumina® (Cat#E7770), following manufacturer´s instructions and 13 cycles of amplification