title: RNASeq of Plasmodium falciparum 3D7 WT compared to MSP2 KO
design: RNASeq of Plasmodium falciparum 3D7 WT compared to MSP2 KO
library_name: WT 1_p
library_construction_protocol: Cultures were tightly synchronised and harvested at late schizont. Cultures were pelleted, resuspended in TriZol (Invitrogen) and incubated for 5 minutes at 37°C before either immediate RNA extraction or freezing at -80°C. Chloroform (Sigma Aldrich) was added, mixed and then spun at 12,000g for 30 minutes at 4°C. Supernatant was collected and mixed with an equal volume of 70% Ethanol and processed using the RNeasy Kit (Qiagen) to extract purified RNA. RNA was DNAse l (Qiagen) treated for 30 mins at room temperature and cleaned up on RNeasy columns according to the manufacturers protocol. Absence of gDNA was checked by qPCR of gDNA and DNAse I treatment repeated if necessary. Library construction and sequencing was performed by Victorian Clinical Genetics Services (VCGS) Australia. Library was prepared using llumina stranded total RNA kit with Ribo -Zero Plus. QC was performed after libary prepration and prior to sequencing.