title: GSM9563971: Immortalized Alveolar type 2 Cell  type 2 Cell line 2; Homo sapiens; RNA-Seq
library_construction_protocol: The immortalized AT2 cell lines were collected and trypsinized after 3–4 passages of 2D culture. The cells were then resuspended in Hanks' balanced salt solution supplemented with 2% fetal bovine serum (FBS), 10 mM HEPES, and 0.1 mM EDTA (HBSS+ buffer). Dead cells were excluded using 7-AAD dye staining. total RNA from the single cells were extracted, reverse transcribed and amplified. Library preparation was performed according to the manufacturer's instructions using the Chromium Single Cell 3' Reagent Kits v2 protocol (10x Genomics, Pleasanton, CA, USA). Briefly, the cell suspension was resuspended in master mix and, together with partitioning oil and Gel Beads, loaded onto Chromium microfluidic chips to generate Gel Beads-in-Emulsion (GEMs). Within each GEM, poly(A) mRNA from individual cell lysates was reverse transcribed into barcoded, full-length cDNA incorporating an Illumina Read 1 primer sequence, a Unique Molecular Identifier (UMI), and a 10x Cell Barcode. The barcoded sequencing libraries were quantified by quantitative PCR using the KAPA Library Quantification Kit for Illumina platforms. Sequencing libraries were loaded onto a NextSeq 500 instrument.