title: GSM6865854: PLC-PRF-5, 25OHD 100nM, 48 h; Homo sapiens; OTHER
library_construction_protocol: MagAttract HMW DNA Kit (Qiagen, Hilden, Germany, 67563) was applied for genomic DNA extraction. To eliminate linear chromosomal DNA and purify circular DNA, the genomic DNA was treated with Plasmid-Safe ATP-dependent DNase (Lucigen) at 37°C, after 6 days of digestion, heat inactivate the exonuclease by incubating at 70°C for 30 minutes. To eliminate mitochondrial DNA, DNAs were eluted and digested with PacI (NEB) for 10 hours, and then extracted with Phenol/chloroform/isoamyl alcohol (PCI) solution (25:24:1). Amplify the remaining circular DNA in the reaction by MDA using φ29 DNA polymerase and random hexamer primers (REPLI-g Mini Kit (Qiagen, Hilden, Germany, 150025)), followed by sample clean up (Agencourt AMPure XP magnetic beads (Beckman Coulter, Brea, CA, USA, A63880)).