GSM6859078: scRNA_E95_mCherry_and_GFP_mCherry_dual; Mus musculus; RNA-Seq
Extraembryonic tissues were removed, and pooled embryos were dissociated using 0,25% Trypsin-EDTA, at 37 ºC for 10 minutes. The cells were washed with 10% FBS/DMEM, then stained with DAPI in 2% FBS/HBSS with 0,5 mM EDTA (FACS buffer) at 4 ºC for 8 minutes. Cells were resuspended in FACS buffer and the indicated populations were sorted into PBS using the BD FACSAria Fusion. The sorted populations were labelled with distinct MULTI-seq surface barcodes, and the cell suspension was filtered with Flowmi Cell Strainers 40 µM before subjected to single-cell RNA sequencing (10x Genomics, Chromium™ Single Cell 3' v3.1). Single-cell RNA-seq libraries were generated following the manual instructions of 10x Genomics, Chromium™ Single Cell 3' v3.1, with the exception of fewer PCR cycles in the cDNA amplification step, where the MULTI-seq surface barcodes were also amplified. MULTI-seq libraries were generated parallel to the single-cell RNA-seq libraries. scRNA-Seq