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Figure 4 | Virology Journal

Figure 4

From: Substitution of adeno-associated virus Rep protein binding and nicking sites with human Chromosome 19 sequences

Figure 4

Site-specific integration at AAVS1. (A) Diagrams (not to scale) of the AAV2 and AAVS1 primer pairs used in nested PCR assays. Each arrow represents a set of two nested primers used in a PCR assay. The number below the AAV2 rep primer set indicates the number of bases from the 5' end of the second primer to the end of the AAV2 genome. The number below the AAVS1 primer set indicates the number of bases from the trs to the 5' end of the second primer in the AAVS1 primer set. (B) Nested PCR assays using the AAVS1 primer set with the AAV2 rep gene primer set. The templates were genomic DNA isolated from HeLa cells infected with the indicated volumes of AAV2-containing supernatants produced using pSub201(-) (wt), pVM108 (108) or pVM113 (113). PCR products were resolved on a 1% agarose gel and stained with ethidium bromide. Single primer controls were done by using only the AAVS1 primer or the AAV2 primer in the second PCR. The template for the single primer controls was DNA from cells infected with 100 μl AAV2. (C) Nested PCR assays using the AAVS1 primer set with the AAV2 ITR primer set. The single primer control used only the AAV2 primer in the second PCR.

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