GSM9347396: Wild type leptotene/zygotene spermatocytes_rep2; Mus musculus; RNA-Seq
Testes were dissected, decapsulated to remove the tunica albuginea, and placed in Dulbecco's phosphate-buffered saline (DPBS) on ice. Then, seminiferous tubules were incubated in digestion buffer containing 120 U/mL of collagenase type I in DPBS at 35 °C for 10 minutes with gentle shaking to dissociate interstitial cells. Tubules were subsequently digested in a second buffer containing 0.25% trypsin and 5mg/mL DNase I at 35 °C for 8 minutes to obtain a single-cell suspension. mRNA reverse transcription, and cDNA amplification were performed using the Discover-sc WTA Kit V2 (Vazyme, N711). cDNA fragmentation and adapter ligation were subsequently carried out using the TruePrep Flexible DNA Library Prep Kit for Illumina (Vazyme, TD504).