GSM6383065: 3m HOX, scRNAseq; Mus musculus; RNA-Seq
Lungs from the experimental C57BL/6 mice were inflated with enzymes, chopped with a razor blade into fine chunks, put into prewarmed enzyme with DNase for digestion, and placed in a 37 °C incubator and rotated for 15min. Then lungs were passed through a 1mL pipet tip. Place the digestion mix back into the incubator and rotate for another 15min. Stop the digestion with FBS and pass the homogenized digested lysate through a 70um filter (Fisher Scientific, Rockford, IL). After centrifugation, incubate with 1X red cell lysis buffer (Sigma-Aldrich, St. Louis, MO) for 3 minutes. After centrifugation and resuspension with Leibovitz media containing 3% FBS, filter lysate with 70um filter, washing with Leibovitz media. Fresh lung single cell digests furnished 1X104 cells, which were loaded on a 10x Genomics v3 scRNA chip using 10x Genomics v3 scRNA reagents. Library preps were done, and completed libraries were sequenced on an Illumina NovaSeq 6000 instrument with 25,000 reads/cell targeted. Raw sequencing data was processed through bcl2fastq2 (Illumina, San Diego, CA), resulting in 1 fastq file per library.