GSM8353852: Endosperm replicate 3; Zea mays; RNA-Seq
10 endosperms from 3 independent ears of the B73 inbred line were manually dissected, transferred into 2mL of LB01 buffer (15mM Tris, 2nM Na2EDTA, 0.5mM spermine, 80mM KCL, 20mM NaCl, 0.1% (v/v) Triton X-100, pH = 8.0, 60U/mL RNasin Plus RNAse Inhibitor (Promega)), on a glass container put on ice. Endosperms were finely chopped with a razor blade during around 30s and the solution was aspired through a 100µm nylon mesh with a needle-less syringa. The filtered solution was stained with 20µg/mL of propidium iodide (PI) during 5min before nuclei were purified by Fluorescence Activated Cell Sorting (FACS) on a BD FACSAria II device. Around 100.000 nuclei were sorted in a 1.5mL Eppendorf and were subsequently centrifuged 5 min at 500g to concentrate them at the bottom of the tube. Nuclear suspensions were loaded into a Chromium Single Cell 3' GEM, Library & Gel Bead Kit (V3 chemistry for repeat 1, V3.1 chemistry for repeat 2+3, 10X Genomics) according to the manufacturer's instructions.