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rcAAV Assay via PCR

PCR and sequencing. The following primers were used for PCR amplification for analysis of rcAAV: D region, 5′-CTCCATCACTAGGGGTTCC-3′ (AAV2 nt 126 to 144); rep, 5′-GGCAGATGCCCGTCAAGGT-3′ (AAV2 nt 379 to 361); and cap, 5′-CAGAGATGTGTACCTTCAG-3′ (AAV2 nt 4026 to 4044).  Episomal DNA was subjected to 35 cycles of PCR amplification using the D-region–rep or D-region–cap primer set. Products were separated on a 1% 3:1 agarose-NuSieve gel and detected by ethidium bromide staining. PCR products were isolated from the gel and cloned into an expression vector for sequence analysis.

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