GSM8207770: Rat 29 [GRC369-29_S6]; Rattus norvegicus; RNA-Seq
The frozen sperm pellet was homogenized in the mirVana Lysis Buffer (ThermoFisher Scientific) in Lysing Matrix D tubes (MP Biomedical) on a Percellys 24 Tissue Homogenizer, followed by a 5-minute incubation at 65◦C. Extraction was continued as described in the mirVana protocol. The concentration of isolated small RNAs (< 200 nucleotides) was determined using Qubit and the quality was assessed using RNA 6000 Nano chips on an Agilent 2100 Bioanalyzer. All samples had RNA integrity number between 2 – 4. Library preparation was performed using the CleanTag Small RNA Library Prep Kit (TriLink Biotechnologies). The pooled library concentrations were determined using the Qubit dsDNA High Sensitivity Assay and quality was assessed using the High Sensitivity DNA kit on the Agilent 2100 Bioanalyzer. Sequencing was performed using the NextSeq 500/550 High Output Kit v2.5 on a NextSeq 500 system set to single end reads at 75 cycles.