alias: GSM4766538
title: GSM4766538: KO-rep1; Plasmodium yoelii yoelii; RNA-Seq
library_construction_protocol: For gametocyte sorting, the gametocytes of the DFsc7 and DFsc7;Δap2-o3 strains were enriched by centrifugation (1,900×g for 10 min) on a 48% Nycodenz solution, and then applied for cell sorting using MoFlo Astrios EQS (Beckman Coulter). Female gametocytes were sorted out through the signal of mCherry. After sorting, the purity of cells was assessed by FACS analysis. Total RNA of parasites was extracted using TRIzol reagent (Invitrogen, #15596026). After extraction of total RNA from purified female gametocytes, mRNA was isolated by Oligo(dT) beads and further enriched by removing rRNA using Ribo-ZeroTM Magnetic Kit (Epicentre, #MRZG126). The enriched mRNA was first fragmented using fragmentation buffer and then reverse transcribed to cDNA with random primers. Second-strand cDNA was synthesized by DNA polymerase I, RNase H, dNTP, and buffer. Then the cDNA fragments were purified with QiaQuick PCR extraction kit, end-repaired, poly(A) added, and ligated with Illumina sequencing adapters. The ligation products were size-selected by agarose gel electrophoresis, PCR amplified, and sequenced using Illumina HiSeqTM 2500 by Gene Denovo Biotechnology (Guangzhou, China).
identifiers.namespace: GEO
identifiers.id: GSM4766538
attributes.tag: GEO Accession
attributes.value: GSM4766538