GSM6360693: w1118 biol rep 1; Drosophila melanogaster; RNA-Seq
200 pairs of testes for each replicate from wild type, cpes mutant and Rescue (Bam-Gal4 and UAS-CPES) Drosophila males were dissected and total RNA was extracted using Trizol and RNA-Clean and concentrater column. About 1ug of RNA in 20ul was submitted for bulk RNAseq using Illumina machine (HWI-ST1276). Total RNA sample quality was measured using Nanodrop (OD260/280), gel electrophoresis and Agilent 2100 to measure RNA integrity. Susequently mRNA was enriched using oligo(dT) beads, fragmented, cDNA was synthesized using random hexamers and reverse transcriptase. After first strand synthesis a custom second strand Synthesis was performed using nick translation method. the final cDNA library was processed via a series of steps including purification, terminal repair, A-tailing, ligation of seequencing adaptors , size selection and PCR enrichment.After quality check libraries were fed into Illumina machines accroding to a activity (>2nM) and expected data volume.