GSM9261628: Testis, WT, bio rep2; Mus musculus; RNA-Seq
Frozen tissues were ground into a fine powder using a mortar and pestle, and RNA was extracted from approximately 20 mg of powdered tissue via the miRNeasy Mini Kit (Qiagen, 217004) per manufacturer's instructions. In brief, tissues were lysed using QiAzol. Chloroform extraction was performed followed by centrifugation at 12,000 x g for 15 minutes at 4oC. The resulting aqueous solution was isolated, and 1.5 volumes of 100% ethanol was added to each sample and transferred to a RNeasy Mini for on-column DNase treatment. Final elution was by 30 uL of RNase-free water. RNA quality was assessed by Agilent TapeStation; samples with RQN > 7.2 were selected for library preparation and sequencing. Libraries were generated from polyA-selected RNA using manufacturer's recommendations for Illumina 2 x 150 basepair sequencing. All samples were sequenced to a minimum depth of 35 million per sample.