GSM6720045: 2-cell embryo - Dam-LMNB1 - CBABL6F1 homozygous (KIN1971_index03); Mus musculus; OTHER
At the time of collection, the zona pellucida was remove using Tyrode's acid (Sigma, T1788-100ML), embryos were washed in M2 medium (Sigma, MR-015-D) and dissociated into single cells by incubating in TryplE (Thermo Fisher, 12605010) for approximately 5 minutes and pipetting up an down with a mouth pipet. Embryos were dissociated one by one and the resulting single cells were placed in M2 medium. They were then transferred to a 384-well plate (one single cell per well) containing 5uL of mineral oil and 100nL of CELseq primer (scDam&T) or 100nL of DamID buffer (0.15% NP40 and 2mM dNTPs) using a mouth pipet. The scDam&T protocol and library preparation was performed as previously described in detail (PMID: 32350457, Supplementary Methods: scDamID2).