GSM9759974: Dam-Lamin B1 (KIN12927.index22); Mus musculus; OTHER
Mothers were sacrificed at E16.5 (48 hours after IUE) and whole brains from the pups were collected for immunostainings in ice-cold Leibovitz's L-15 medium (Gibco, 11415064), washed in PBS and fixed by immersion in 4% paraformaldehyde (PFA, Sigma, 30525-89-4) in PBS at 4 °C overnight. The scDam&T-seq processing was largely performed as previously described in {Markodimitraki 2020}, but reaction volumes of all mixes were equimolarly halved (0.5x) to reduce overall processing costs. As such, the added volumes per well were 50 nl CEL-seq2 primer (before sorting, with 5 µl mineral oil), 50 nl lysis mix, 75 nl reverse transcriptase mix, 925 nl second strand mix, 250 nl proteinase K mix, 150 nl DpnI mix, 50 nl of DamID adapter (1 µM concentration, 25 nM final during ligation) and 450 nl scDam&T T4 ligase mix. The overall composition of the mixes remains unaltered and the incubation times and temperatures are as described.