GSM8967233: Testis, WT, PD13; Mus musculus; RNA-Seq
Single-cell suspensions were prepared from PD13 mouse testis tissue following the Chromium Single Cell 3' Solution protocol (V3 chemistry). The testis samples were washed twice with cold 1× phosphate-buffered saline (PBS). Cell viability was assessed using a Haemocytometer (Thermo Fisher Scientific). Cell concentration was then measured using Countstar and adjusted to 1000 cells/μl. For samples with concentrations below the required threshold (less than 400 cells/μl), cells were pelleted and resuspended in a reduced volume, followed by recounting to confirm the new concentration. The single-cell testis suspension was loaded into Chromium microfluidic chips with 3' V3 chemistry and barcoded using a 10× Chromium Controller (10X Genomics). RNA from the barcoded testis cells underwent reverse transcription, and sequencing libraries were constructed using reagents from a Chromium Single Cell 3' V3 reagent kit (10X Genomics) following the manufacturer's protocol. Sequencing was performed on Illumina NovaSeq according to the manufacturer's instructions.