HepAD38 cccDNA ExoI/III-PCR
To induce HBV replication and cccDNA formation in HepAD38 cells, tetracycline (tet) was withdrawn from the culture medium, and cells were cultured for 18 days before harvest. The harvested cells were subjected to Hirt DNA extraction, followed by exonuclease I and III (Exo I/III) (NEB, #M0293 and #M0206) digestion. The treated DNA samples were subsequently column-purified and amplified by PCR using cccDNA-specific primers (S6 Table) and Phusion High-Fidelity DNA Polymerase (Thermo Fisher Scientific, #F530S). The PCR products were further purified by QIAquick PCR Purification Kit (Qiagen, #28106) prior to sequencing.