GSM3899354: YC3B; Cryptococcus neoformans var. grubii H99; RNA-Seq
- Fix and freeze yeast culture: - Pre-chill a 50mL tube with 6mL 100% Methanol  on dry ice (tightly capped so CO2 does not dissolve in MetOH). - Add 10mL of culture to the chilled MetOH, cap tube, shake to mix, place on dry ice for up to 5 min. - Spin 2 min at ~2500g, pour off supernatant. - Resuspend cell pellet in 1mL ice-cold H2O and transfer to 2mL screw-cap tube. - Spin 1min at 5000g, remove supernatant carefully, and freeze pellet in sealed tube at -80C. - Freeze-dry the pellet in a lyophilizer until dry. - Extract RNA using the RNAeasy plant and fungi kit (Qiagen) RNAtagseq essentially as described in Shishkin et al., Nat. Methods 2015, "Simultaneous generation of many RNA-seq libraries in a single reaction", doi: 10.1038/nmeth.3313. Edits to protocol at https://github.com/ewallace/protocols/blob/master/RNAtagseq.md