RNA-Seq of M. pachydermatis ATCC 14522 grown in Sabouraud Dextrose Medium
M. pachydermatis ATCC 14522 (CBS 1879) was grown on Sabouraud Dextrose Agar for 3 days at 35C. Ten individual colonies were picked and used to inoculate 10 ml of liquid Sabouraud Dextrose medium in a 50-ml Erlenmeyer flask. The flask was incubated at 30C and 130 rpm shaking for 48 hours. Ten ml of fresh growth medium were placed into a new 50-ml flask and inoculated with 1 ml from the starter culture. The flask was incubated at 35C and 200 rpm shaking for 3 hours. Cells were collected by centrifugation, flash frozen in dry ice ethanol, and stored at -80C until RNA was extracted. RNA was extracted using a hot phenol method. Genomic DNA was removed by DNase I treatment and RNA purified using a QIAgen RNeasy kit. Paired-end RNA-Seq libraries were prepared with the KAPA Hyper Stranded mRNA Library Kit (Roche); 250-bp reads were collected.