GSM8000798: Normal Rabbit Serum E11.5 TrkC Cut&Run rep4; Mus musculus; OTHER
DRG were dissected from E11.5 BacE-GFP expressing embryos (Appel et al 2016) and dissociated with Trypsin B/Collagenase H.  GFP positive cells were FACS isolated and 300,000-600,000 cells were used for Cut&Run analysis with anti-Brn3a (1:200) (kindly provided by E.Turner), anti-Islet1 (1:50) (Abcam 20670), home made anti-Runx3 (1:200), anti-H3K27Ac (1:50) (Abcam 4729) antibodies or Normal Rabbit Serum (NRS, 1:200). GFP positive TrkC neurons were bound to ConA beads, permeabilized, incubated with anti-Brn3a, anti-Islet1, anti-Runx3, anti-H3K27Ac or NRS and targeted by MNase cleavege. Antibody bound DNA was extracted by phenol chloroform. For ATAC analysis 50,000 cells were used. The cells were lysed and the DNA was exposed to the transposition reaction. Library was prepared using NEBNEXT Ultra II DNA Library Prep Kit for Illumina (#E7645). DNA fragments were PCR amplified with Illumina primers for no more than 15 cycles and library fragments up to ~250 bp (insert plus adaptor and PCR primer sequences) were SPRI (x0.8) purified.