GSM5695948: NF54; Plasmodium falciparum; RNA-Seq
Total RNA from saponin-lysed parasites was extracted using Trizol (Invitrogen) and Direct-Zol RNA MiniPrep Plus Kit (Zymo Research) rRNA was removed from Total RNA using Illumina Ribo Zero Gold for human/mouse/rat kit. Using Illumina TruSeq RNA Sample Library Preparation v2 kit (Illumina, San Diego, CA), Messenger RNA was fragmented into small pieces using divalent cations under elevated temperature.The cleaved RNA fragments were copied into first strand cDNA using reverse transcriptase and random primers. Second strand cDNA synthesis followed, using DNA Polymerase I and RNase H. The cDNA fragments then went through an end repair process, the addition of a single 'A' base, and then ligation of the adapters. The products were then purified and enriched with PCR to create the final cDNA library. Libraries were pooled and sequenced on Illumina HiSeq4000 sequencer with single-end 50 cycles. rRNA was removed from Total RNA using Illumina Ribo Zero Gold for human/mouse/rat kit. Using Illumina TruSeq RNA Sample Library Preparation v2 kit (Illumina, San Diego, CA), Messenger RNA was fragmented into small pieces using divalent cations RNA-seq