Reactivation of the silenced X-chromosome during mouse germline development
Reactivation of the silenced X-chromosome during mouse germline development
Experimental F1 embryos were generated by crossing Xist+/∆ (B6) dams carrying Oct4-EGFP transgene, and CAST sires, producing F1 female (XX(Xist∆)) and males (XY) that were analyzed.   Following timed natural matings, F1 embryos were harvested daily at E10.5 to E16.5. Gonads were dissected and dissociated into single-cell suspension. EGFP fluorescence was used to isolate the fluorescently marked germ cells with FACS. Viable individual germ cells and somatic cells were sorted by FACS into 96-well plates containing 4µl lysis buffer. Please see 'nucleic acid library construction protocol'. Single-cell RNA-seq libraries were generated as described in Picelli et al., 2014, Nature Protocols. Full-length RNA-seq from single cells using SMART-seq2.