PCR-based in situ lysis gel analysis of HHV-8 genomes during initial infection.

Cells were lysed in the wells of an agarose gel at the indicated times, and extrachromosomal DNA that had been separated by electrophoresis was extracted from sequential slices of the gel and analyzed by PCR amplification of a 394-bp fragment from the HHV-8 T1.1 gene. (A) At left is the configuration of HHV-8 genomes in latently infected BCBL-1 cells before and after lytic cycle induction with 3 mM sodium butyrate for 72 hours. At right is the configuration of HHV-8 before (top), at 24 hours after (middle), and at 72 hours after (bottom) primary infection in fMVDECs with HHV-8. (B) The configuration of HHV-8 genomes 8 hours after infection with HHV-8 in fMVDECs in an independent experiment. The positions of sequential PCR-amplified DNAs relative to the top and bottom of the original in situ lysis gel are indicated from left to right. The relative positions of circular and linear forms are also indicated.