RNAseq of chytrid infected P. agardhii, ball phenotype
Total RNA was extracted using the Qiagen RNeasy PowerWater Kit (Qiagen, Germantown, MD, USA) by first removing the filter from the cartridge under sterile conditions in a laminar flow hood, then following the manufacturers instructions with the following notes: 60 L of Qiagen RNase Free DNase solution was incubated for 30 minutes at room temperature to elute the RNA from the extraction column. Remaining DNA was removed using the Ambion Turbo DNA-free kit (Thermo Fisher Scientific) according to manufacturers instructions. Extracted RNA was sent to Discovery Life Sciences (Huntsville, AL), where the samples were treated to reduce rRNA using TruSeq Stranded Total RNA with Ribo-Zero Plant kit (Illumina, San Diego, CA). RNA was sequenced on an NovaSeq 6000 with paired-end reads of 150 base pairs.