Fluorescence Activated Nuclei Sorting.

(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.