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Testes were harvested from 6-week-old male C57 mice from the control and Ara-C-treated groups and immediately processed to generate single-cell suspensions. Briefly, testicular tissues were finely minced and enzymatically digested to dissociate cells. The digested tissues were filtered through a 40-um cell strainer to obtain a uniform single-cell suspension. After centrifugation and washing, cells were resuspended in PBS containing 0.04% BSA. Cell number and viability were determined before loading onto the single-cell platform. Single-cell 3' RNA-seq libraries were prepared using the 10x Genomics Chromium platform following the manufacturer's instructions. Individual cells were partitioned into GEMs with barcoded beads, enabling transcript labeling with cell-specific barcodes and UMIs during reverse transcription. After cDNA amplification, sequencing libraries were prepared and sequenced on the Illumina NovaSeq 6000 platform. |
RNA-Seq |
TRANSCRIPTOMIC SINGLE CELL |
RANDOM |
PAIRED
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