HBV-HepG2-NTCP cccDNA heat/PSAD-PCR
HBV infection of HepG2-NTCP (HBV-HepG2-NTCP) was 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 digestion. The treated DNA samples were subsequently column-purified and amplified by PCR using cccDNA-specific primers 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.