title: GSM8001459: testis at differentiating stage; Sebastes melanops; RNA-Seq
library_construction_protocol: the tunica albuginea of the testis was removed, and the testis was incubated in 10 ml Leibovitz-15 medium (Gibco Invitrogen, Grand Island, NY) containing 20 mg/ml collagenase H (Roche Diagnostics, Mannheim, Germany), 0.25% trypsin (Worthing- ton Biochemical, Lakewood, NJ), 5% fetal bovine serum (Gibco), and 0.05% DNase I (Roche Diagnostics) for 4 hours at 28℃. Cell suspensions were fertilized through 40 μm strainers. Following three rounds of centrifugation at 200 rpm, and discarding the supernatant after each centrifugation, the testicular cells were suspended in 1ml PBS. cells were diluted following manufacturer's instructions, and 33.8 μl of total mixture buffer along with cells were loaded into the 10x Chromium Controller using Chromium Single Cell 3′ v2 reagents.