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1 March 2006 Heated Oligonucleotide Ligation Assay (HOLA): An Affordable Single Nucleotide Polymorphism Assay
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Abstract

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.

W. C. Black, N. Gorrochotegui-escalante, and N. M. Duteau "Heated Oligonucleotide Ligation Assay (HOLA): An Affordable Single Nucleotide Polymorphism Assay," Journal of Medical Entomology 43(2), 238-247, (1 March 2006). https://doi.org/10.1603/0022-2585(2006)043[0238:HOLAHA]2.0.CO;2
Received: 14 July 2005; Accepted: 12 September 2005; Published: 1 March 2006
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