GSM8388879: P. knowlesi in blood, Nycodenz, centrifuge; Plasmodium knowlesi; RNA-Seq
Resuspended enriched iRBC pellets were further diluted in PBS-BSA to a concentration of 60,000 cells/mL using a Scepter 3.0 cell counter (Merck). 1mL of cell suspension was loaded into each HIVE to settle into the pico-wells via gravity or using a centrifuge according to modified manufacturer's directions. The incubation time for the HIVE loading was increased from 30 minutes to 1 hour at room temperature (gravity) and from 3 to 5 minutes (centrifuge). HIVEs were stored at -20°C until library preparation to mimic using a -20°C freezer in the field. Library preparation was conducted following manufacturer's directions with 10 extra PCR cycles added to the whole transcriptome amplification (WTA). All samples were sequenced by on a NovaSeq6000 (Broad Institute, USA) according to the manufacturer's recommendations. Sequencing data has been deposited in the NCBI sequencing read archive.