GSM6217981: L37_Richter's_PBMC_GEX; Homo sapiens; RNA-Seq
Cells were washed and partitioned into nanoliter-scale Gel Bead-In-EMulsions (GEMs), where all generated cDNA share a common 10x Genomics Barcode but uses a pool of ~750,000 barcodes to separately index each cell's transcriptome. The silane magnetic beads and Solid Phase Reversible Immobilization beads were used to clean up the GEM reaction mixture and the barcoded cDNA was then amplified in a PCR step. Libraries were prepared using the Chromium Next GEM Single Cell 5' Kit (v2) (PN-1000263): Library Construction kit (PN-1000190), Chromium Next GEM Chip K Single Cell Kit (PN-1000286), Chromium Single Cell Human BCR Amplification Kit (PN-1000253), and Dual Index Kit TT Set A (PN-1000215) (10x Genomics). Optimal cDNA amplicon size was achieved using Covaris machine prior to library construction. The P7 and R2 primers were added during the GEM incubation and the P5, and R1 during library construction via end repair, A-tailing, adapter ligation and PCR. The final libraries contain the P5 and P7 primers used in Illumina bridge amplification.