title: GSM6733517: C934-KO; Mus musculus; RNA-Seq
library_name: GSM6733517
library_construction_protocol: The 'tween with salts and tris' (TST) method (Drokhlyansky et al., 2019; Slyper et al., 2020) was used to isolate nuclei.  A pair of mouse whole adrenals were placed into a well of a 6-well plate on ice with 1 ml of TST buffer (146 mM NaCl, 10 mM Tris-HCl pH 7.5, 21 mM MgCl2, 1 mM CaCl2, 0.03% Tween-20, 0.01% BSA in nuclease-free water). Tissue was then chopped using scissors for 10 min. The homogenized solution was then filtered through a 70 µm Falcon cell strainer, and additional 1 ml TST buffer was used to wash the cell strainer. Additional TST buffer was added to bring the total volume up to 5 ml. Sample was transferred into a 15 ml conical tube and centrifuged at 500g for 5 min at 4 °C in a swinging bucket centrifuge, and the pellet was suspended in ST buffer (146 mM NaCl, 10 mM Tris-HCl pH 7.5, 21 mM MgCl2, 1 mM CaCl2 in nuclease-free water). The nuclei solution was filtered through a 35 µm Falcon cell strainer twice, stained with 4′9,6-diamidino-2-phenylindole (DAPI) and directly sorted using Flow Activated Nuclei/Cell Sorting with a FACSAriaII instrument. Library was performed according to the manufacter's instructions (single cell 3' v3 protocol, 10x Genomics).