Journal article

A novel detection method for heteroduplex DNA using carbodiimide-induced interrupted primer extension

T Tabone, R Cotton, N Mathew, J Des Parkin, J Savige

Journal of Nucleic Acids Investigation | Published : 2010


Direct sequencing may be problematic in demonstrating mutations where inherited disease results from multiple different heterozygous variants in large genes. We describe here a novel mutation screening method based on the ability of carbodiimide to bind mismatched DNA and interrupt primer extension thereby identifying both a heterozygous variant and its location. This assay detected all four classes of DNA mismatch in 550 bp engineered plasmid fragments and in two dominantly inherited renal diseases. In patients with thin basement membrane nephropathy, the method demonstrated multiple variants within a single amplicon including some close to the primer binding site. This method also detected..

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