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. 2020 Oct 16;31(19-20):1054–1067. doi: 10.1089/hum.2020.118

Figure 1.

Figure 1.

Replication assay of wtAAV and rAAV following cotransfection of wt and rAAV plasmids detected by Southern blot analysis. (A) Schematic maps of the wtAAV2 and rAAV-CBA-Luc, including the probes used for detection. Southern blot analysis of wtAAV2 (B) or rAAV DNA replication (C) using the indicated probes. Equimolar of total plasmids transfected into 293 cells. To assess wtAAV DNA replication, cells were transfected with pxx680/pSSV9/pcDNA3.1; for rAAV, cells were transfected with pxx680/pXR2/pAAV-CBA-Luc; for wtAAV+rAAV group, cells were transfected with pxx680/pSSV9/pAAV-CBA-Luc; for control, cells were transfected with pAAV-CBA-Luc/pXR2/pcDNA3.1. Hirt DNA was recovered at 48 h post-transfection. Equal amounts of DNA was digested with Dpn I, separated on 0.8% agarose gel, and subjected to Southern blot analysis using 32p-labeled rep or CBA probe. CBA, Cytomegalovirus enhancer/chicken β-actin promoter; ITR, inverted terminal repeats;. rAAV, recombinant adeno-associated viral; wtAAV, wild-type AAV.