GSM5903105: SGET Day 6 mPGCLC rep1; Mus musculus; ncRNA-Seq
mPGCs were isolated by microdissection of pooled pairs of male genital ridges away from mesonephros. These were treated in trypsin for 20 min at 37C to achieve single isolates. Cells were then incubated with surface marker antibodies SSEA1-660 and CD61-PE and FACS sorted directly into Qiazol. mPGCLCs were isolated from Day 6 trypsin-dossiciated Embryoid Bodies using reporter gating (Stella-GFP positive and Esg1-tdTomato negative) via FACS and sorted directly into Qiazol. Small RNA fractions was isolated using the Qiagen miRNAeasy-micro kit. Small RNA libraries were made using the Perkin-Elmer NextFlex small RNA Kit v3, following the instructions for low-input.These contain 4 random Ns at the adapter ends.