title: GSM8993129: Dam-P2A control for ATF4 DamID, replicate1; Mus musculus; OTHER
library_construction_protocol: Genomic DNA was extracted from cells using a QiaAmp DNA Micro kit (Qiagen 56304) according to the manufacturer's instructions. gDNA was fragmented using DpnI digestion overnight at 37°C, followed by dsAdR adaptor ligation using T4 DNA ligase for 2 h at 16°C, followed by ligase inactivation for 10 min at 65°C. Digestion using DpnII was then performed for 2 h at 37°C, followed by DpnII inactivation for 20 min at 65°C. The reaction mixture was subsequently purified using Seramag magnetic beads, followed by PCR amplification using the DreamTaq protocol. The processed DamID fragments (AlwI-digested and sonicated) were  purified using Seramag magnetic beads and  Illumina sequencing libraries were then constructed using NEBNext Ultra II DNA Library Prep Kit. After a further Seramag bead purification, the adaptor-ligated DNA was amplified for 6 cycles using NEBNext Ultra II Q5 Master Mix with i5 and i7 index primers to prepare the libraries.