Analysis of wild type and GAT201 deletion mutant strain reactivation in RPMI supplemented with and without 10% heat-inactivated fetal calf serum at 37°C.

We wanted to find out the transcriptomic response of growth arrested wild type (KN99a) and Gat201 deletion mutant Cryptococcus yeast cells reactivating in cell culture media with and without serum. Overall design: 56-sample RNA-seq timecourse of reactivation of growth-arrested 2 x wild type (KN99a + KN99a) and 2x Gat201 deletion mutant (Madhani and Bahn deletion collection) Cryptococcus neoformans in two different cell culture media at 37°C. Two biological samples of cells were pre-grown for 5 days in 20mL YPD at 30°C, 200rpm. A 2ml sample of this was taken as the "0 minute" control timepoint. We then inoculated 3.5ml in 2 different cultures of 100mL pre-warmed media: (RPMI, RPMI+Serum) at 37°C and grew for 30, 120 or 240 min. 15ml of culture was collected at each time point and fixed in 33& ice cold methanol, spun down astored at -80°C.