GSM8565559: CT_1 (biol rep 1); Sporisorium scitamineum; RNA-Seq
RNA extraction was performed with RNeasy Plant Mini Kit (Qiagen), with the protocol modified only for the lysis step. This step involved the addition of 500 µL of glass beads and 450 µL of RLC buffer with β-mercaptoethanol, followed by three lysis cycles of vortex and ice (30 seconds each). RNA integrity was verified with the Agilent RNA 6000 Nano kit on the 2100 Bioanalyzer (Agilent Technologies). RNA quality and quantification was performed using the Qubit™ RNA High Sensitivity – HS kit (Thermo Fischer Scientific) and Qubit 2.0 equipment (Thermo Fischer Scientific) Twelve libraries were prepared using the TruSeq Stranded mRNA Sample Prep LS Kit (Illumina) following the manufacturer's protocol, and the quality of the libraries was evaluated using the 2100 Bioanalyzer with the Agilent DNA 1000 Kit, following the manufacturer's protocol. Libraries were quantified using the KAPA qPCR Master Mix kit (Roche) on the Viia7 Real-Time PCR System equipment (Applied Biosystems) and were sequenced on the Illumina HiSeq 2500.