GSM6359048: PDL1, scRNAseq; Mus musculus; RNA-Seq
GSM6359048
Single cell suspension in RT Master Mix was loaded on the Single Cell Chip and partition with a pool of about 750,000 barcoded gel beads to form nanoliter-scale Gel Beads-In-Emulsions (GEMs). Upon dissolution of the Gel Beads in a GEM, the primers with the unique cell barcodes were released and mixed with cell lysate and Master Mix. Incubation of the GEMs then produced barcoded, full-length cDNA from poly-adenylated mRNA. Silane magnetic beads were used to remove leftover biochemical reagents and primers from the post GEM reaction mixture. Full-length, barcoded cDNA was amplified by PCR to generate sufficient mass for library construction. Enzymatic Fragmentation and Size Selection are then used to optimize the cDNA amplicon size prior to library construction. The final libraries contained the P5 and P7 primers used in Illumina bridge amplification. Analysis steps such as demultiplexing, alignment, and gene counting, and visualization to generate expression data with single-cell resolution were performed.