GSM8342824: scRNA_lung_shNKX2-1_Vehicle; Mus musculus; RNA-Seq
Single-cell RNA sequencing libraries were generated by using a Chromium Controller instrument (10x Genomics, Pleasanton, CA, USA) and Chromium Single Cell 3' Reagent Kits v2 according to the manufacturer's instructions. The library preparation was conducted in accordance with the manufacturer's guidelines using the single-cell 3' v2 protocol from 10x Genomics. In summary, cells were suspended in a master mix and combined with partitioning oil and gel beads in a chip to create gel bead-in-emulsion (GEM) structures. The poly-A RNA from each GEM was reverse transcribed into cDNA, incorporating an Illumina R1 primer sequence, a Unique Molecular Identifier (UMI), and a 10x Barcode. The pooled barcoded cDNA was purified using Silane DynaBeads, PCR amplified, and fragments of appropriate sizes were selected using SPRIselect reagent for subsequent library construction. During library construction, the Illumina R2 primer sequence, paired-end constructs with P5 and P7 sequences, and a sample index were introduced.