PFNF54-Pfs16-GFP-LUC gametocyte time course from commitment to maturity

Plasmodium falciparum gametocytes undergo 14 days of gametocytogenesis to generate mature, transmissible gametocytes. This timecourse delineates the transcriptional profiles of gametocytes from commitment to maturity. P. falciparum NF54-Pfs16-GFP-LUC parasites were induced to produce gametocytes by culturing in glucose-deficient media , before removing asexual parasites from the population by daily 5% D-sorbitol treatments from day 1-4 and thereafter culturing the parasites in glucose positive media and grown under stationary conditions for the entire time course. The timecourse, resulting in a total of 16 samples was hybridized against a Cy3-labeled reference pool of 3D7 mixed stage asexual parasites and NF54-pfs16-GFP-LUC mixed stage gametocytes on a two-color array. Starting at 2 days before induction, samples were Samples were harvested from culture by 0.01%(w/v) saponin addition on days -2 to 7 and via density centrifugation using Nycoprep 1.077 cushions on days 8-13 and stored at -80˚C until RNA isolation using TRIZOL reagent along with on-column extraction using the Qiagen RNeasy kit (Qiagen, Germany).