GSM2545654: wt/cl 2; Drosophila melanogaster; RNA-Seq
RNA was extracted using the RNeasy mini spin kit (Qiagen) and stored at -80°C. Purification of poly-A containing mRNA is performed by using poly-T oligo-attached magnetic beads. The mRNA is then fragmented into small pieces using divalent cations under elevated temperature. The cleaved RNA fragments are copied into first strand cDNA using reverse transcriptase and random primers. Subsequently, a second cDNA strand is synthesized using DNA Polymerase I and RNase H. Then, a single 'A' base and the adapter are added to the cDNA fragments. Finally, the products are purified and enriched with PCR to create the final cDNA library. Illumina TruSeq Stranded mRNA library prep.