title: GSM9013756: Phox2b H2B-CFP low intensity 2; Mus musculus; ATAC-seq
library_construction_protocol: Cold-active protease (Native Bacillus Licheniformis) with added DNaseI were used to dissociate pooled E16.5 Phox2b H2B-CFP guts (stomach to anus, 6 guts for 4828-MS-15+4828-MS-16 and 12 pooled guts for 4828-MS-17+4828-MS-18) at 5°C in a waterbath and thermo-mixer. Dissociated cells were pelleted and washed in L-15 media with added BSA, HEPES, penicillin, streptomycin, amphotericin, and DNaseI. Cell suspensions were filled through 40µm nylon mesh. Fluorescence-activated cell sorting was used to simuntaneously sort CFP high intensity and CFP low intensity cells from nonfluorescent cells. Nuclei were isolated using the 10x Genomics protocol number CG000209_Rev D. Prior to encapsulation, nuclei were counted via Countess Automated Cell Counter or hemocytometer to determine concentration of nuclei. These nuclei were encapsulated using the 10x Genomics platform for snATAC-seq on bright nuclei and dim nuclei separately. Nuclei were encapsulated using the 10x Genomics platform for snATAC-seq on bright nuclei and dim nuclei separately. Two replicates were generated with the first bright and dim replicates generated from 6 pooled fetal guts and the second generated from 12 pooled fetal guts.