RNA-seq of Plasmodium falciparum: trophozoite stage, cytoplasmic fraction
Messenger RNA was purified from total cytoplasmic and nuclear RNA samples using NEBNext Poly(A) nRNA Magnetic Isolation module (NEB; E7490S) with manufacturers instructions. Once mRNA was isolated, strand-specific RNA-seq libraries were prepared using NEBNext Ultra Directional RNA Library Prep Kit for Illumina (NEB; E7420S). Libraries were amplified for a total of 12 PCR cycles (45 s at 98C followed by 15 cycles of [15 s at 98C, 30 s at 55C, 30 s at 62C], 5 min 62C).