I am working on OBI among HIV patients naive to HAART. I expect to detect HBV DNA in HIV positive clients -ve for HBsAg but +ve for HBcAb using nPCR. My target regions are the BCP/PC, Core region and the Small S-region of the HBV genome. The primers used are EP1-1 and EP1-2, EP2-1 and EP2-2 for the BCP/PC region. HB7F and HBAS6, CB3 and HBAS5 for the core region. HB1F and HS4R, B2 and HS4R for the S-region. In running the amplicons on gel, no bands are seen even with my positive control. All protocols were duly followed. What could be the possible cause? Is it advisable i digest the genomic DNA with an enzyme though I am interested in performing RFLP and cloning?

Thanks