GSM3615474: triptolide_1; Mus musculus; ncRNA-Seq
Total RNAs were extracted from the testes using TaKaRa MiniBEST Universal RNA Extraction Kit (TaKaRa, Dalian, China) according to the manufacturer's protocol with some modifications. Briefly, on-column DNase I digestion was performed to eliminate trace amounts of DNA contamination for all samples. Finally, the concentration and purity of the total RNA samples were evaluated using a NanoPhotometer (IMPLEN, Munich, Germany) with optical density measurements at 1.8 < A260/A280 < 2.2. The integrity of RNA was always checked by electrophoresis in 1.5% agarose gels and the 2100 Bioanalyzer (Agilent Corporation, Palo Alto, CA, USA). After the rRNA was depleted from total RNA using Ribo-Zero™ rRNA removal Kit (Illumina, San Diego, CA, USA), next generation sequencing library preparations were constructed according to the manufacturer's protocol (NEBNext® Ultra™ Directional RNA Library Prep Kit for Illumina®). Sequencing was carried out using a 2 x 150 paired-end (PE) configuration. The sequences were processed and analyzed by GENEWIZ, Inc. (Suzhou, China).