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. 2017 Mar 22;7:91. doi: 10.3389/fcimb.2017.00091

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Copyright © 2017 Li, Sheng, Wang, Yang, Liang, Huang, Liu, Li, Yang, Yang, Jia, Xie, Wang, Hao, Du, Xu, Zhou, Li, Sun, Tong, Li, Qiu and Song.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Analysis of integrated HBV DNA in the stable HBV cell line A64. (A) Integrated HBV DNA in the stable HBV A64 cell line and the gRNA target sites in the repeat region of the 1.1 HBV genome copy. (B) The 4,049-bp DNA fragment representing the 3,362-bp integrated HBV DNA (1.1 copies) plus a 687-bp pTriexHBV1.1-derived flanking sequence was efficiently amplified from cellular genomic DNA using the integrated HBV-specific primers P1 and P2. (C) PCR analysis using the A1AT and HBV S-gene primer sets conducted on total genomic DNA and circular duplex DNA to assess the effect of extraction on circular duplex DNA. (D) PCR analysis of the integrated HBV DNA or circular duplex DNA isolated from cells using the P1/P3 and S-gene primer sets. Using P1 and the HBV core region-specific primer P3, the HBV S-gene amplicons were predicted to be 542- and 572-bp, respectively. Primers P1/P3 did not amplify circular duplex DNA.