GSM3882388: Matched_D2T0940_Rep2; Plasmodium chabaudi chabaudi; ssRNA-seq
Samples were collected every three hours, over a period of 30 hours, from 09:00 GMT on day 4 to 15:00 GMT on day 5 post infection. This corresponds to a starting time of ZT1.5 for matched, and ZT13.5 for mismatched infections. At each sampling point, four mice from each group (matched/mismatched) were sacrificed and thin blood smears and RBC counts (via flow cytometry; Beckman Coulter) were taken by tail bleeds, and 50µl of blood was taken via cardiac puncture (added to 200µl of RNAlater and frozen at -80C) for RNAseq analysis. RNAlater was removed from the samples preserved with RNAlater through centrifugation and 1 ml of Trizol was added to lyse the samples. 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 11 timepoints were anlyzed further
GEO Accession
GSM3882388