GSM8425440: 20170221_WT_ES; Mus musculus; RNA-Seq
For male germ cells isolation from testis, a protocol adapted from the work of Bastos et al. (2005)was used, as described in Brouze et al. (2024). Total RNA from testes and collected spermatides was isolated with TRI Reagent (Sigma) following manufacturer's protocol. Tissues were homogenizedin TRI Reagent using glass Dounce homogenizer while collected cell pellets were suspended in TRI Reagent and lysed by pipetting. DNA traces from final RNA samples were removed using TURBO DNA free kit (Thermo Fisher Scientific) before further experiments. rRNAs were removed from total DNA-free RNA samples with Ribo-Zero Gold rRNA-removal Kit (Illumina). rRNA-free samples were cleaned-up by 3M sodium acetate precitipation method. KAPA Stranded RNA-Seq Library Preparation Kit (KAPA Biosystems) was used for sequencing libraries preparation, with random primers used for cDNA synthesis step. On each step of the procedure samples were cleaned-up using AMPure XP Reagent magnetic beads. Quality and fragment size distribution of the libraries was determined by electrophoresis on Agilent 2100 Bioanalyzer (Agilent Technologies Inc.).