GSM8462834: Pair 2, Replicate 1, GEX; Mus musculus; RNA-Seq
Sequencing was performed with four 10-week-old males from a single litter; two Tex14m/m and two wild-type animals. Testes were dissected and two testes from a single animal were pooled. Testes were dissociated and cells were isolated as described previously (Green et al. 2018). Single-cell libraries were generated using the 10x Genomics Chromium Controller and following the manufacturer's protocol for 3' v3.1 chemistry with feature barcoding technology for cell multiplexing (10x Genomics). In brief, cells from each animal were counted and tagged with unique cell multiplexing oligo barcodes. Labelled cells from one wild type and one mutant were pooled equally and processed for cell encapsulation targeting 12,000 cells (~6,000 cells per mouse), followed by library preparation. This was done in duplicate to capture ~12,000 cells per mouse. Labelled cells from the second wild type and second mutant were pooled and processed similarly. Final library quality was assessed using the LabChip GXII HT (PerkinElmer) and libraries were quantified by Qubit (ThermoFisher). Pooled libraries were subjected to 28x10x10x151 bp paired-end sequencing according to the manufacturer's protocol (Illumina NovaSeq XPlus) to generate ~25,000 reads per cell.