GSM9179486: NMR_lung_animal#510, rep 2; Heterocephalus glaber; RNA-Seq
Lungs were inflated with 1X PBS and collected from NMR and mice. Tumors were removed from normal tissue manually using forceps and scissors. Tumors were dissociated into single cells as described in the supplementary methods. Cell suspensions were washed twice with 1X PBS containing 0.04% BSA. Single-cell RNA sequencing was conducted using the 10X Genomics Chromium X/IX (10X Genomics, Pleasanton, CA) at the Molecular Genomics Core of the Moffitt Cancer Center and Research Institute. Following wash steps, cells were resuspended in the same buffer (1X PBS with 0.04% BSA) according to the 10X Genomics cell preparation protocol. Cell viability and counts were assessed using AO/PI dual fluorescent staining and analyzed with the Nexcelom Cellometer K2 (Nexcelom Bioscience LLC, Lawrence, MA). Subsequently, cells were loaded onto the 10X Genomics Chromium Single Cell Controller at a concentration of 1,000 cells/µl, aiming to encapsulate 10,000 cells per sample. Individual cells, reagents, and 10X Genomics gel beads were encapsulated into nanoliter-sized Gel Beads in Emulsion (GEMs). Reverse transcription of poly-adenylated mRNA occurred within each droplet at 53°C. cDNA libraries were prepared in a single bulk reaction following the 10X Genomics Chromium NextGEM Single Cell 3' Reagent Kit v3.1 guidelines, and 50,000 sequencing reads per cell were generated using the Illumina NovaSeq 6000 instrument
GEO
GSM9179486_r1