GSM4968864: Fetal gonads, sample 1; Bos taurus; RNA-Seq
Slow-frozen gonads were thawed at 37°C in a water bath,  rinsed in ice-cold PBS and minced into ~0.1 mm pieces. Minced tissue was then incubated in Collagenase Type IV (800U/mL) (LS004186, Worthington) supplemented with DNase I (6U/mL) (04536282001, Roche Applied Science) at 37°C in a prewarmed orbital shaker for 10 min at 150 rpm. After dissociation, the cell suspension was filtered and dead cells were removed by magnetic bead sorting using a Dead Cell Removal Kit (13009010, Miltenyi) following the manufacturer's protocol. Viable cells resuspended in 0.4% BSA were submitted on ice to the UC Davis DNA Technologies & Expression Analysis Core for library preparation and sequencing. Cells were captured using a 10x Chromium controller targeting 10,000 cells per sample. Libraries were prepared using the Chromium Single Cell 3' v2 chemistry for sample 1 and v3 chemistry for sample 2 according to the manufacturer's protocol.