Instrument: Illumina HiSeq 2500
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: PBMCs were isolated from whole blood using density centrifugation. Cells were then processed by the 10X Genomics Chromium single cell platform using the manufacturer's 3' protocol to obtain a pooled library. The 10x Genomics Chromium machine partitions cells into Gel Bead-In-EMulsions (GEMs), where all generated cDNA share a common 10x Genomics barcode but uses a pool of about 750,000 barcodes to separately index each cell's transcriptome. The P7 and R2 primers were added during the GEM incubation and the P5, and R1 during library construction via end repair, A-tailing, adaptor ligation and PCR. The final libraries contained the P5 and P7 primers used in Illumina bridge amplification. The library was prepared for sequencing using an Illumina 150 cycle (v2 chemistry) high output kit and sequenced on an Illumina HiSeq 2500 to obtain paired-end reads.