title: GSM3398455: RPE NoTx_C1; Homo sapiens; OTHER
library_construction_protocol: RPE cells were lysed with gentle extraction buffer prepared in 1X PBS containing 1% v/v Triton X-100 (Sigma-Aldrich) and 1 mM EDTA for 15 min on ice. Lysate was centrifuged at 1000Xg for 10 min at 4 °C to pellet-out nuclei. The lysate supernatant was used as the cytoplasmic fraction. Pippin-fractionated ssDNA samples were converted to dsDNA by the Seq Plex Enhanced DNA Amplification Kit (Sigma-Aldrich, SEQXE) without additional fragmentation to enrich for DNA between 200-800 bp. Amplification was monitored by RT-PCR, and cycle number (20–25 cycles) was set as 2-3 cycles after the amplification plateau, as suggested by the manufacturer. 1 μg of dsDNA library was prepared for sequencing with the NEXTflex® Rapid DNA Sequencing Kit (Bioo Scientific).