GSM7780242: cotyledon seed, replicate 1, scRNA-Seq; Glycine max; RNA-Seq
Fresh cotyledon stage seeds were hand chopped in lysis buffer (15mM Tris pH7.5, 20mM NaCl, 80mM KCl, 0.5mM Spermine, 5mM 2-ME, 0.05% Triton X-100, 80U/ml RNase inhibitor (Roche, 3335399001)), then filtered through 40um and 20um nylon mesh.  Nuclei were diluted with 3 volumes cold wash buffer (20mM MES pH5.4, 0.4M Mannitol, 20mM KCl, 0.1% BSA, 80U/ml RNase inhibitor), and resuspended in 1ml cold wash buffer. DAPI-stained nuclei were counted on a hemocytometer, spun again and resuspended to achieve a target concentration of 3500 nuclei/ul. Library was constructed using 10,000 nuclei, following 10X Genomics protocol for Chromium Next GEM Single Cell Multiome ATAC+Gene expression v1 assay.