GSM3102840: MBC10.shWDR5.Rep3; Homo sapiens; RNA-Seq
2.5x10^4 cells are lysed and processed according to TRUSeq RNA Sample Prep Kit Illumina protocol. Briefly, after cell lysis the poly-A containing mRNA molecules were purified using oligo-dT attached magnetic beads. Following purification, the mRNA is 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. Second strand cDNA synthesis follows, using DNA Polymerase I and RNase H. The cDNA fragments then go through an end repair process, the addition of a single 'A' base, and then ligation of the adapters. The products are then purified and enriched with PCR to create the final cDNA library for HiSeq 2000 Illumina sequencing.