Fig. 4

Validation of RNA processing pipeline on malaria-infected clinical samples from TRAC2 study. a Giemsa smear estimated parasitaemia and total RNA yields obtained from 200 µl of iRBCs from ten patients' blood samples. b, c A heatmap showing correlation of whole transcriptome amplified from patients' blood samples to 3D7 IDC reference transcriptome based on b FPKM values obtained from RNA sequencing, c spotted oligo microarray hybridization. The highest PCC values are shown in numerals. The same total RNA input of 250 ng was used for reverse transcription of all samples. All ten patients' samples in panel a, b, and c have been selected from various clinical sites across South East Asia and sorted from the highest to the lowest parasitaemia. d A scatterplot depicting positive correlation between 18S-Pf/18S-Hs peak heights ratio and percentage of P. falciparum unique reads. All data were obtained from Bioanalyzer electropherograms and RNA sequencing of 171 TRAC2 clinical blood samples. Blue and red dotted lines indicate minimum values of the parasite reads proportion (16%) and peak ratio (0.4), respectively, in order to obtain 3 million unique parasite reads when multiplexing 20 samples on Illumina HiSeq 4000 lane.