Heated oligonucleotide ligation assay (HOLA): an affordable single nucleotide polymorphism assay
Autor(es): Black, W. C.; Gorrochotegui-Escalante, N.; Duteau, N. M.
Resumo: 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 approximately US$1.25 with two-allele SNPs or approximately 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 alpha -glycerophosphate dehydrogenase locus in the mosquito Anopheles gambiae s.s.
Palavras-Chave: Genotypes; Aquatic insects; Hybridization; Entomology; Dehydrogenases; Nucleotides; Denaturation; Trypsin; Single-nucleotide polymorphism; Assays; Oligonucleotides; dehydrogenase; Aedes aegypti; Culicidae; Anopheles gambiae
Imprenta: Journal of Medical Entomology, v. 43, n. 2, p. 238-247, 2006.
Descritores: Aedes aegypti - Immunology
Data de publicação: 2006
