title: GSM8584250: DamID-seq, E7.75 embryo, Dam-Lhx1, rep 2; Mus musculus; OTHER
library_construction_protocol: Genomic DNA was extracted with the Quick-gDNA MicroPrep (ZymoResearch) kit. Extracted gDNA was digested with DpnI and DamID adapters were then ligated using T4 DNA Ligase. After ligation, the DNA was digested with DpnII and the DamID fragments of interest were amplified with the KAPA HiFi Polymerase. Following PCR amplification, the samples were cleaned up using Wizard SV Gel and PCR Clean-Up System (Promega) and eluted into 90 µL dH20 + 10 µL CutSmart buffer (NEB). Samples were sonicated with a Bioruptor Plus (Diagenode) for 5 minutes, high power, 30s on/30s off. 10% of the sample was ran on a 0.8% gel to check fragment sizes then the DNA was digested with AlwI overnight at 37°C. Following clean-up with AMPure XP beads, 500 ng of each sample was used for Library preparation with NEBNext Ultra II DNA Library Prep Kit for Illumina (NEB).
identifiers.namespace: GEO
identifiers.id: GSM8584250_r1