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. 2020 Jun 12;31(11-12):664–678. doi: 10.1089/hum.2019.277

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© Karen Guerin et al., 2020; Published by Mary Ann Liebert, Inc.

This Open Access article is distributed under the terms of the Creative Commons Attribution Noncommercial License (http://creativecommons.org/licenses/by-nc/4.0/) which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and the source are cited.

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Figure 2. — Analysis of DNA conformation of rAAV vectors. (A) DNA extracted from the rAAV vectors pAAV-hSyn-EGFP (AAV-50465) and pAAV-CAG-GFP (AAV-37825) were left untreated (Ctrl) or incubated in the presence (37°C + SacII) or absence (37°C) of the restriction enzyme SacII and products separated by agarose gel electrophoresis. ΦX174 DNA was incubated in the presence of SacII, HaeIII or left untreated. The table lists the size of the viral genomes and the expected products after digestion. (B) Undigested and SacII digested DNA from plasmid DNA or extracted vDNA was amplified with primers targeting the region surrounding the SacII cleavage site or gfp as a control. SacII amplification was normalized to gfp amplification, and fold change of undigested to SacII digested samples was calculated and plotted. gfp, green fluorescent protein gene; rAAV, recombinant adeno-associated virus; vDNA, virus DNA. Color images are available online.