title: GSM7744881: benign_1, snATAC-seq; Homo sapiens; ATAC-seq
library_construction_protocol: Isolation procedure was adapted from Nuclei Isolation from Complex Tissues for Single Cell Multiome ATAC + Gene Expression Sequencing (CG000375). Nuclei were isolated from a 50mg piece of frozen tissue. The tissue was cut into 5 small sections with a razor blade. The tissue pieces were homogenized for 1.5 minutes with a tissue homogenizer and pestle in 10mM Tris-HCl, 10mM NaCl, 3mM MgCl2, 0.1% Nonidet P40 Substitute, and 1mM DTT in nuclease free water. The tissue was allowed to dissociate in the lysis buffer for 5 minutes before passing through a 70mm cell strainer. The solution was then centrifuged for 5 minutes at 500 x g at 4°C. The supernatant was removed and replaced with 1 mL 1% BSA in DPBS and allowed to incubate for 5 minutes before resuspension. The nuclei were centrifuged again at 500 x g, 4°C for 5 minutes. The pellet was resuspended in 1mL 1% BSA in DPBS and passed through a 30mm cell strainer. All steps were performed on ice. Nuclei were stained with 7-Aminoactinomycin D (7-AAD) Ready Made Solution 1:100 (Sigma, SML-1633) and sorted with a FACSAria III Cell Sorter.  Nuclei were collected into 10% BSA. Prepared per manufacturer's instructions (10X Genomics, 1000176; CG000209 Rev F)