GSM9736868: Testis tissue, control mouse, replicate 3; Mus musculus; RNA-Seq
Total RNA was extracted from mouse testis tissues using TRIzol Reagent according to the manufacturer's instructions. RNA quality and integrity were detected by NanoDrop 2000 and agarose gel electrophoresis, and RNA integrity was evaluated using Agilent 5300 Bioanalyser. Only RNA samples with concentration ≥20 ng/μL, total RNA >1 μg and RQN >4.5 were used for subsequent library construction. RNA-seq libraries were constructed following Illumina® Stranded mRNA Prep, Ligation protocol at Shanghai Majorbio Bio-pharm Biotechnology Co., Ltd. Briefly, poly(A) mRNA was enriched by oligo(dT) magnetic beads and fragmented into about 300 bp fragments. First-strand and second-strand cDNA were synthesized sequentially, followed by end repair, A-tailing and adapter ligation. Target fragments were selected and amplified by PCR. Qualified libraries were quantified by Qubit 4.0 and sequenced on NovaSeq X Plus platform with PE150 mode.