title: GSM8686555: IPF lung sample 1, snRNA-seq; Homo sapiens; RNA-Seq
library_name: GSM8686555
library_construction_protocol: Frozen lung tissues were collected from the most basal segments of the right lower lobes, and carefully excised to obtain approximately 50 mg of tissue free of visible airway structures, vessels, mucin, or blood clots. Tissues were immediately placed into a nuclei isolation buffer (20 mM Tris-HCl pH 7.5, 292 mM NaCl, 2 mM CaCl2, 42 mM MgCl2, 1% CHAPS, and 2% BSA) and homogenized using a Dounce homogenizer to mechanically dissociate nuclei from cells. The resulting homogenate was filtered through a 40 µm strainer, and nuclei were pelleted by centrifugation at 300 × g for 2 minutes at 4°C. Nuclei were resuspended in 0.04% BSA buffer to a final concentration of 1,000 nuclei/µl. Single-nuclei RNA sequencing (snRNA-seq) libraries were prepared using the 10x Genomics 5' Kit v2, following the manufacturer's protocol (10X Genomics). Libraries were sequenced on an Illumina NextSeq 550 platform using 75-cycle paired-end sequencing.