GSM9856929: luad1; Homo sapiens; RNA-Seq
Single-cell suspensions were loaded onto the 10x Genomics Chromium Controller to generate Gel Bead-in-Emulsions (GEMs). Within each individual GEM, single cells were lysed, and the released intracellular polyadenylated mRNA transcripts were captured by the barcoded oligonucleotides present on the 10x Gel Beads. Reverse transcription was subsequently performed within the intact GEMs to synthesize single-cell barcoded full-length cDNA. Following reverse transcription, the emulsion was disrupted, and the uniquely barcoded cDNA was pooled and amplified via polymerase chain reaction (PCR). The amplified cDNA was purified using SPRIselect beads. The purified cDNA then underwent enzymatic fragmentation, end repair, and A-tailing. Finally, sequencing adapters and sample-specific indices were incorporated via PCR to construct the final single-cell RNA-seq libraries, adhering strictly to the manufacturer's protocol for the 10x Genomics Chromium Single Cell 3' Reagent Kits.