title: GSM2367699: wild-type head rep1; Drosophila melanogaster; RNA-Seq
library_construction_protocol: kmg ctrl, kmg KD, dMi-2 ctrl, and dMi-2 KD testes without accessory glands were dissected in PBS for 30 minutes and immediately frozen in liquid nitrogen before transferred to -80°C. Wild-type heads were manually decapitated from the body and frozen and stored as in testes preparation. Total RNA was extracted using TRIzol (Invitrogen).  RNA was further purified by RNeasy mini column (Qiagen) to minimize residual salts. RNA quality was assessed by checking rRNA integrity using Bioanalyzer. ~30ug of total RNA was used to purify polyA RNA by using Poly(A)Purist MAG Kit (Thermo Fisher, Cat#AM1922). 100ng of purified mRNA was used to make a strand-specific sequencing library by dUTP incorporation method using KAPA Stranded RNA-Seq Library Preparation Kit Illumina® Platforms (KAPA Biosystems, Cat#: KK8400) following manufacturer’s instructions. All RNA-Seq libraries were sequenced by NextSeq Mid Output mode 150bp paired end. Between 3 to 4 samples were pooled in a single lane, yielding on average 48 million reads per sample (34 to 65 million). All sequencing was performed by the Stanford Functional Genomics Facility (SFGF).
attributes.tag: GEO Accession
attributes.value: GSM2367699