GSM6432741: E14.5 Fev-YFP+ sorted; Mus musculus; ATAC-seq
Nuclei were isolated from sorted WT or YFP+ cells. Cells were centrifuged at 500 rcf for 5 min at 4C. All the supernatant was removed without disrupting the cell pellet. 100 ul chilled 0.1x lysis buffer was then added and pipetted 5 times to fully mix the buffer with the cells, and incubated for 3 min on ice to achieve full cell lysis. 1ml chilled wash buffer was added to the lysed cells to terminate the lysis. The lysed cells were centrifuged at 500 rcf for 5 min at 4C and supernatant was gently removed. Nuclei were then resuspended in 50ul wash buffer and transferred to a 200ul tube. Nuclei were spun down at 500 rcf for 5 minutes and resuspended in 10ul 1x Nuclei buffer (10x Genomics, PN 2000153). 2 ul of the nuclei suspension was used for determining the concentration of nuclei using hemacytometer, and the cell nuclei quality was assessed at the same time. 12,500 nuclei for the whole pancreas sample, 5,000 nuclei for the eFev-YFP+ sample and 28,500 for the E17.5 whole pancreas were then immediately processed with Chromium Next GEM Single Cell ATAC Reagent Kits v1.1. Isolated nuclei were transposed and partitioned using the Chromium Next GEM Single Cell ATAC Kit v1.1, following manufacturer's instructions. The resulting library was then run on the Illumina NovaSeq 6000 platform.