GSM3884641: PchSR10KO_D3T1130_rep1; Plasmodium chabaudi chabaudi; ssRNA-seq
Total RNA was isolated from TRIzol treated samples according to the manufacturer's instructions (Life Technologies). Strand-specific mRNA libraries were prepared from total RNA using TruSeq Stranded mRNA Sample Prep Kit LS (Illumina) according to the manufacturer's instructions. Briefly, at least 100ng of total RNA was used as starting material to prepare the libraries. PolyA+ mRNA molecules were purified from total RNA using oligo-T attached magnetic beads. First strand synthesis was performed using random primers followed by second strand synthesis where dUTP were incorporated in place of dTTP to achieve strand-specificity. Ends of the double stranded cDNA molecules were adaptor ligated and the libraries amplified by PCR for 15 cycles. Libraries were sequenced on Illumina HiSeq 4000 platform with paired-end 100/150bp read chemistry according to manufacturer's instructions. Data from 14 timepoints were anlyzed further excluding the first three time points.econd strand synthesis where dUTP were incorporated in place of dTTP to achieve strand-specificity. Ends of the double stranded cDNA molecules were adaptor ligated and the libraries amplified by PCR for 15 cycles. Libraries were sequenced on Illumina HiSeq 4000 platform with paired-end 100/150bp read chemistry according to manufacturer's instructions. Data from 14 timepoints were anlyzed further