Journal Article

Heated Oligonucleotide Ligation Assay (HOLA): An Affordable Single Nucleotide Polymorphism Assay

in Journal of Medical Entomology

Published on behalf of Entomological Society of America

Volume 43, issue 2, pages 238-247
Published in print March 2006 | ISSN: 0022-2585
Published online October 2014 | e-ISSN: 1938-2928 | DOI: http://dx.doi.org/10.1093/jmedent/43.2.238

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Most single nucleotide polymorphism (SNP) detection requires expensive equipment and reagents. The oligonucleotide ligation assay (OLA) is an inexpensive SNP assay that detects ligation between a biotinylated "allele-specific detector" and a 3′ fluorescein-labeled "reporter" oligonucleotide. No ligation occurs unless the 3′ detector nucleotide is complementary to the SNP nucleotide. The original OLA used chemical denaturation and neutralization. Heated OLA (HOLA) instead uses a thermal stable ligase and cycles of denaturing and hybridization for ligation and SNP detection. The cost per genotype is ≈US$1.25 with two-allele SNPs or ≈US$1.75 with three-allele SNPs. We illustrate the development of HOLA for SNP detection in the Early Trypsin and Abundant Trypsin loci in the mosquito Aedes aegypti (L.) and at the α-glycerophosphate dehydrogenase locus in the mosquito Anopheles gambiae s.s.

Keywords: single nucleotide polymorphisms; oligonucleotide ligation assay; mosquitoes

Journal Article.  4229 words.  Illustrated.

Subjects: Disease Ecology and Epidemiology ; Entomology

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