GSM5025570: miP 1 (RNA-seq); Drosophila melanogaster; RNA-Seq
Secondary cells were isolated from dissected male accessory glands.  Briefly, glands were dissected and treated with 1X TrypLE in Schnieider's media in the presence of 1'%fetal bovine serum and 1% Pen-Strep.  The glands were then titurated to separate the cells and sorted by FACs.  RNA was then isolated from purified cells using the Epicentre Masterpure RNA purification kit. 2ng RNAs from either sorted SCs or TRAP experiments were used to generate cDNAs using polydT primers for reverse transcription and SMARTer technology for amplification (Takara). Libraries for sequencing were prepared using Nextera XT kit (Illumina), and were sequenced using multiplexed, single reads of 100 nucleotides (20-30Million reads per sample).