Preparation and Validation of PCR-generated Positive Controls for Diagnostic Dot Blotting

Abstract
Allele-specific oligonucleotides (ASOs) are small, single-stranded nucleotide polymers (∼18–20 bases in length) of diagnostic utility given their ability to hybridize to single-stranded DNA target molecules in a sequence-specific, temperature-dependent manner. Accordingly, two target molecules differing in composition by a single base can be distinguished by hybridization with an ASO that is complementary to one of the target molecules but noncomplementary to the other by a single base. The latter target produces a single basepair-mismatched double strand that dissociates at a lower washing temperature than the fully complementary, double-stranded molecule.