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. 2016 Mar 30;5:16019. doi: 10.1038/mtm.2016.19

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Copyright © 2016 Official journal of the American Society of Gene & Cell Therapy

This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/4.0/

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Design of AAV5 inverted terminal repeats (ITR) quantitative polymerase chain reaction (qPCR). (a) Top: 5’ ITR secondary hairpin structure of wild-type AAV5 and localization of the AAV5 ITR qPCR-specific primers and probe. Bottom: Corresponding 71-bp PCR product. (b) ITR5 qPCR using pAAVPGK plasmid linearized with ScaI, or digested with KpnI to create ITRs with free ends. The differences between mean quantification cycles (C_q) obtained in each experimental condition with an equal amount of plasmid quantified by spectrophotometry are shown. (c) r-square, slope, and intersection values of the qPCR tests shown in a. (d) Titration of an rAAV5 vector sample using the two different linearized plasmids (ITR5 versus free-ITR5). The mean and standard deviations listed for rAAV5 vectors are from four different dilutions of the samples. *P < 0.05.