RNA-Seq of Mus musculus:WT testis SG rep3
The ribosomes of Ribosome RPL39L+/+ and Ribosome RPL39L-/- mouse testis were separated by 20%-50% sucrose gradient ultracentrifugation, and 80S monosome, 2-6 and 7+ polysome components were obtained.Spermatogonia were isolated from 6 days post-partum (dpp) mice, and round and elongated spermatids from adult mice (56-72 dpp) using the STA-PUT method, based on sedimentation velocity at unit gravity. Spermatocytes were isolated from adult mice (56-72 dpp) using the STA-PUT method or using fluorescence-activated cell sorting (FACS).The RNA was extracted using the Trizol reagent kit (Invitrogen) with the quality checked by Agilent 2100 Bioanalyzer (Agilent Technologies) and agarose gel electrophoresis. After enrichment using Oligo (dT) beads, the RNA was fragmented, and reverse transcripted into cDNA with random primers, which were purified by QiaQuick PCR extraction kit (Qiagen). After end repair, poly(A) addition, and ligation to Illumina sequencing adapters, the ligation products were size selected by agarose gel electrophoresis, PCR amplified and sequenced using HiSeq2500 (Illumina) by Gene Denovo Biotechnology Co. (Guangzhou, China).