GSM5208275: T20061; Plasmodium falciparum; RNA-Seq
Total RNA was extracted using Trizol and ZYMO DirectZol96 extraction kit following protocol described in Kucharski, M., Tripathi, J., Nayak, S. et al. A comprehensive RNA handling and transcriptomics guide for high-throughput processing of Plasmodium blood-stage samples. Malar J 19, 363 (2020). Reverse transcription and cDNA synthesis was done following modifed Smart-Seq2 protocol using only oligo(d)T primers to enrich for mRNA transcripts (Kucharski, M., Tripathi, J., Nayak, S. et al. A comprehensive RNA handling and transcriptomics guide for high-throughput processing of Plasmodium blood-stage samples. Malar J 19, 363 (2020)). Purified amplicon (cDNA) was used to generate sequencing libraries using Illumina Nextera XT kit as described in manufacturer's protocol. Libraries were analyzed on the Bioanalyser High-Sensitivity DNA chips (Agilent, Cat. No.: 5067-4626), subsequently pooled (24 samples per lane) and sequenced on Illumina HiSeq4000 platform generating 150 bp paired end reads with 110 Gb data output generated per lane.