HBV-PHH cccDNA heat/PSAD-PCR
HBV infection of PHH (HBV-PHH) cells were conducted and the cells were infected for 6 days before harvest. The harvested cells were subjected to Hirt DNA extraction, followed by heat-denaturation and PSAD treatment. 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.