GSM7774049: 220819L06_exp220519_MS63_SCGE-15nM-BE_polyA-RNA-seq; Saccharomyces cerevisiae; Nakaseomyces glabratus; RNA-Seq
S. cerevisiae and C. glabrata cells were separately pelleted and snap-frozen in liquid nitrogen. Pellets were thawed in ice-cold PBS and mixed. 30 µl of cells in PBS were added to 300 µl TRI Reagent (Zymo Research) and lysed by bead beating using a Fastprep 24. Cell debris was pelleted (14k rpm, 1 minute) and the supernatant recovered. RNA was then extracted using the direct-zol RNA microprep kit (Zymo Research). mRNA was enriched using the NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB, E7490). Indexed sequencing libraries were generated using the NEBNext Ultra II RNA Library Prep Kit for Illumina (NEB, #E7775), pooled, and then sequenced by paired-end (2x150bp) Illumina sequencing. spike-in normalised polyA RNA-seq