GSM8999638: 5FU, Candida, Antibiotics, Sample 3; Mus musculus; RNA-Seq
RNA isolation from Tongue tissues was carried out using the RNeasy Kit (Qiagen) as follows: tongue samples were homogenized in 800µl of RLT buffer containing 1% βeta-mercaptoethanol (Biorad) using a tissue homogenizer. Then, 800µl of Phenol:chloroform:isoamyl-alcohol (Invitrogen) was added to the homogenate in 2 ml screw-capped tubes. Samples were centrifuged, and the aqueous phase was collected. An equal volume of 70% molecular-grade ethanol was added. Kit instructions were followed for subsequent column separation steps. DNAse treatment was performed using the TURBO DNA-free kit (Invitrogen). RNA was analyzed for quality using the Experion automated gel electrophoretic system (Biorad) For the RNA sequencing, cDNA libraries were prepared using the TruSeq RNA kit (Illumina) and subjected to poly(A) enrichment according to the kit protocol. cDNA fragments (100 nucleotides from each end) were sequenced using the Illumina NextSeq platform.