title: GSM8072525: piwi,ovary,rep3; Drosophila melanogaster; RNA-Seq
library_construction_protocol: Total RNA from young wildtype (w1118) and piwi1/piwi2 mutant fly ovaries were extracted, purified following standard phenol-chloroform extraction steps. CAGE library preparation and sequencing were performed by DNAFORM. Qualities of total RNA were assessed by Bioanalyzer (Agilent) to ensure that RIN (RNA integrity number) is over 7.0. The cDNAs were synthesized from total RNA using random primers. The ribose diols in the 5' cap structures of RNAs were oxidized, and then biotinylated. The biotinylated RNA/cDNAs were selected by streptavidin beads (cap-trapping). After RNA digestion by RNaseONE/H and adaptor ligation to both ends of cDNA, double-stranded cDNA libraries (CAGE libraries) were constructed. CAGE libraries were sequenced using single end reads of 75nt on a NextSeq 500 instrument.