GSM5126517: yw replicate #2; Drosophila melanogaster; RNA-Seq
For each replicate, ~100 pairs of ovaries were hand dissected from newly eclosed females. RNA was extracted using TRIzol (Invitrogen) followed by purification on RNeasy spin columns (Qiagen). mRNA is enriched using oligo(dT) beads. For library construction, the mRNA is fragmented randomly by adding fragmentation buffer, then the cDNA is synthesized by using mRNA template and random hexamers primer, after which a  custom second-strand synthesis buffer (Illumina) , dNTPs, RNase H and DNA polymerase I are added to initiate the second-strand synthesis. Second, after a series of terminal repair,  A ligation and sequencing adaptor ligation, the double-stranded cDNA library is completed through size selection and PCR enrichment.