GSM8161056: FEL1-MT1,scRNAseq; Mus musculus; RNA-Seq
Freshly prepared lung cells are FACS purified from mouse chimeric lung generated from blastocyst complementation. The tissues were digested with liberate to prepare single cell suspension. Library was constructed according to the manufacter's instructions (single cell 3' v3.1 protocol, 10X genomics). The cells prepared were resuspended in the the Chromium Single Cell 3' master mix solution and processed by microfluidic partitioning to capture single cells and create next-generation sequencing (NGS) cDNA libraries in Gel Beads in emulsion.The single cells were lysed, and the Gel Bead is dissolved to release barcoded RT oligonucleotides into solution, and reverse transcription of polyadenylated mRNA occurs. This results in all cDNAs from a single cell having the same barcode for mapping back to their original cell of origin. Next, Illumina primer and adapter sequences were incorporated to make paired-ended constructs with P5 and P7 to assist next generation sequencing.