Plasmodium falciparum steady-state mRNA Gametocyte stage
Total RNA was extracted from parasite cultures using Trizol LS Reagent (Life Technologies), followed DNase I (Life Technologies) treatment and mRNA isolation using the GenElute mRNA Miniprep Kit (Sigma-Aldrich). After first and second-strand cDNA synthesis, sequence libraries were prepared using the Encore Multiplexing System (NuGen) according to the manufacturer’s instructions, with the following modifications for the high AT-content of the P. falciparum genome: the libraries were amplified for a total of 15 PCR cycles (5 cycles of [15s at 98*C, 30s at 55*C, 30s at 62*C] followed by 10 cycles of [15s at 98*C, 30s at 63*C, 30s at 72*C]) using KAPA HiFi HotStart Ready Mix (Kapa Biosystems, Woburn, MA).