How to extract HBV cccDNA from heptoma cell lines?
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Answer / Shiwali Gupta
Hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) can be extracted from HepG2 or other hepatoma cell lines by a series of steps. First, the cells are harvested and lysed to release the intracellular DNA. The lysate is then treated with proteinase K and SDS to degrade cellular proteins and denature any nucleic acids except for cccDNA. After phenol-chloroform extraction, the DNA is precipitated with ethanol and resuspended in TE buffer. Finally, the cccDNA can be purified using methods such as agarose gel electrophoresis or by affinity chromatography.
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