GSM8830702: MMTV-PyMT Prrx1 +/+ Tumors Rep3; Mus musculus; ATAC-seq
Mammary gland carcinomas from 14- to 15-week-old MMTV-PyMT mice were manually minced and homogenized in Solution D (20 mM Tris-HCl pH 7.5, 0.1% Tween 20, 0.25 M Sucrose, 25 mM KCl, 5 mM MgCl2). The lysate was filtered (70 μm, 40 μm), centrifuged, and resuspended in Wash Buffer. Nuclei were isolated using Optiprep gradient centrifugation and lysed in 0.1X lysis buffer. After filtration (20 μm) and final centrifugation, nuclei were resuspended in Nuclei Dilution Buffer for downstream processing. Viability and count were assessed using Trypan Blue and an automated cell counter. Single-nuclei suspensions from four mice were processed using the Chromium Next GEM Single Cell ATAC Kit v2 (10x Genomics). Nuclei were transposed, encapsulated with the 10x Chromium Controller, and multiplexed using the Chromium Single Index Kit N. Libraries were sequenced on an Illumina NextSeq 2000 (50 bp read length).