RNA-seq profiles of Madurella mycetomatis mm55 cultured pathogen

Mycetoma is a neglected, chronic granulomatous infection of the subcutaneous tissue, most often caused by the fungal pathogen Madurella mycetomatis. Characteristic of the infection is the formation of grains. Overall design: M. mycetomatis genome strain MM55 was cultured on Sabouraud Dextrose Agar for two to three weeks at 37°C. Mycelium was harvested, sonicated at 28 micron (Soniprep 150, Beun de Ronde, the Netherlands) and incubated at 37°C in RPMI-media supplemented with 0.35 g/L L-glutamine, 1.98 mM 4-morpholinepropanesulfonic acid (MOPS) and 100 mg/L chloramphenicol. After two weeks, the mycelia were separated and washed by vacuum filtration (Nalgene, Abcoude, the Netherlands using a 22 micron filter (Whatman). The mycelial biomass was scraped from the filter, the wet weight was determined and a suspension was prepared containing 100 mg mycelial biomass per ml phosphate-buffered saline (PBS). The suspension was sonicated at 28 micron for 2 minutes (Soniprep 150, Beun de Ronde, the Netherlands), the resulting homogeneous suspension was washed once in PBS and again diluted to a concentration of 4 mg per 40 µL PBS, corresponding to 600-850 CFU/larvae.