title: Illumina HiSeq 2000 sequencing; Differentially expressed genes upon melatonin treatment on Plasmodium falciparum infected red blood cells
design: Differentially expressed genes upon melatonin treatment on Plasmodium falciparum infected red blood cells
library_construction_protocol: Plasmodium falciparum parasite lines (3D7, Pfpk7- and PfpK7 complement clones) were cultured in RPMI 1640 (Invitrogen) supplemented with 37.5 mM HEPES, 7mM D-glucose, 6mM NOH, 25 /mL gentamicin sulphate, 2mM L-glutamine and 10% human serum) and maintained in human erythrocytes under a gas mixture of 5% O2, 5% CO2, and 90% N2 (31). Cultures were synchronized by using 5% sorbitol (32). For Pfpk7- and PfpK7 complement parasite strains, the same medium was supplemented with 2.5 μg/mL blasticidin. Melatonin was added to synchronised parasite cultures (30h p.i) to a final concentration of 100 nM and incubated for 5 hours and 17 hours at 37°C. 'Total RNA was extracted using Trizol reagent (Invitrogen) according to the manufacturer''s instructions. RNA was quantified using the Nanodrop ND-1000 UV/Vis spectrophotometer and RNA integrity was checked by electrophoresis using an Agilent 2100 Bioanalyzer. Total RNA (6000 ng) from each sample was stabilized in RNAStable (Biomatrica) and sent to Scripps Research Institute for HiSeq Illumina sequencing.'
attributes.tag: Experimental Factor: none
attributes.value: compound
attributes.tag: Experimental Factor: Pfpk7 Complement
attributes.value: genotype