Fluorescence Activated Nuclei Sorting.
figureposted on 2021-05-11, 17:27 authored by Daniel Conde, Paolo M. Triozzi, Kelly M. Balmant, Andria L. Doty, Mariza Miranda, Anthony Boullosa, Henry W. Schmidt, Wendell J. Pereira, Christopher Dervinis, Matias Kirst
(A, B) To minimize the contamination of organelles such as chloroplast and remove the debris that could clog the 10× Genomics microfluidic chips, we sort the nuclei using FANS technology using a BD FACSAria™ IIU/III upgraded cell sorter. (C) An initial gate was drawn on a FSC and SSC plot to eliminate the majority of non-nuclear debris and organelles. The final histogram was used to isolate the desired nuclei. 40,000 DAPI+ nuclei were sorted with a total recovery volume of 67–70 μl, into a 1.5 ml RNase free non-stick Eppendorf low binding tube containing 10 μl of NIB WASH.