GSM8776810: VitC scDam&T-seq Dam-LMNB1 (VAN10206.index30); Mus musculus; OTHER
MESCs were fixed at -20C using 70% Ethanol and permeablised using 0.05% Tween. ProteinA-MN was targeted using antibodies against IgG, Lamin B1, H3, H3K4me1, H3K4me3, H3K9me3, H3K27me3 or H3K36me3 and cells were FACS sorted into 384 well plates. MN was activated thorugh 2mM CaCl2 for 30min at 4C at inactivated by EGTA addition. Cells were lysed (protK + NP40), fragments end-repaired (Klenow) and adenine methylated sites were cut (DpnI). Dam and ChIC fragemnts are ligated to blunt end adaptors containing a cell barcode and unique molecular identifier and a T7 promoter. After pooling DNA is amplified by in vitro transcription, before rundom hexamer based reverse transcription and final indexing PCR for sequencing.