GSM1909368: Mutant_Suz12_FRT2A; Drosophila melanogaster; RNA-Seq
RNA-Seq libraries were constructed with the TruSeq RNA sample preparation (Low throughput protocol) kit from Illumina One microgram of total RNA was used for the construction of the libraries. The RNA is fragmented into small pieces using divalent cations under elevated temperature. The cleaved RNA fragments are copied into first strand cDNA using SuperScript II reverse transcriptase and random hexamer primers. The Second strand cDNA is synthesized. These 20 cDNA fragments then go through an end repair process, the addition of a single 'A' base and the subsequent ligation of the adapter. The products are then purified and enriched with 15 cycles of PCR. The final cDNA libraries are validated with a DNA 1000 Labchip on a Bioanalyzer (Agilent) and quantified with a KAPA qPCR kit. For one sequencing lane of a flowcell V3, three libraries were pooled in equal proportions, denatured with NaOH and diluted to 7 pM in hybridization buffer. Cluster formation, primer hybridisation and single read 50 cycles sequencing were performed