Article
Systemic errors in quantitative polymerase chain reaction titration of self-complementary adeno-associated viral vectors and improved alternative methods.
Human gene therapy methods - 1 Feb 2012
Fagone Paolo, Wright J Fraser, Nathwani Amit C, Nienhuis Arthur W, Davidoff Andrew M, Gray John T
Abstract excerpt
Self-complementary AAV (scAAV) vector genomes contain a covalently closed hairpin derived from a mutated inverted terminal repeat that connects the two monomer single-stranded genomes into a head-to-head or tail-to-tail dimer. We found that during quantitative PCR (qPCR) this structure inhibits the amplification of proximal amplicons and causes the systemic underreporting of copy number by as much as 10-fold. We...
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