16,500 cells were run on the 10X Chromium to partition single cells into droplets containing uniquely barcoded beads and processed for sequencing library preparation using the Chromium Single Cell 5â NextGEM v2 chemistry. Amplified full-length cDNA from poly-adenylated mRNA is used to generate 5Ê¹ Gene Expression library. Enzymatic fragmentation and size selection are used to optimize the cDNA amplicon size prior to 5â gene expression library construction. P5, P7, i5 and i7 sample indexes, and Illumina R2 sequence (read 2 primer sequence) are added via End Repair, A-tailing, Adaptor Ligation, and Sample Index PCR. The final libraries contain the P5 and P7 priming sites used in Illumina sequencers.
UMIGS-Maryland-Genomics Illumina Construction Protocol Details