New separation-free assay technique for SNPs using two-photon excitation fluorometry
AUTOR(ES)
Vaarno, Jonne
FONTE
Oxford University Press
RESUMO
A new separation-free method for detection of single nucleotide polymorphisms (SNPs) is described. The method is based on the single base extension principle, fluorescently labeled dideoxy nucleotides and two-photon fluorescence excitation technology, known as ArcDia™ TPX technology. In this assay technique, template-directed single base extension is carried out for primers which have been immobilized on polymer microparticles. Depending on the sequence of the template DNA, the primers are extended either with a labeled or with a non-labeled nucleotide. The genotype of the sample is determined on the basis of two-photon excited fluorescence of individual microparticles. The effect of various assay condition parameters on the performance of the assay method is studied. The performance of the new assay method is demonstrated by genotyping the SNPs of human individuals using double-stranded PCR amplicons as samples. The results show that the new SNP assay method provides sensitivity and reliability comparable to the state-of-the-art SNaPshot™ assay method. Applicability of the new method in routine laboratory use is discussed with respect to alternative assay techniques.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=506825Documentos Relacionados
- Two-photon excitation microscopy of tryptophan-containing proteins
- Simultaneous two-photon excitation of distinct labels for dual-color fluorescence crosscorrelation analysis
- A protease assay for two-photon crosscorrelation and FRET analysis based solely on fluorescent proteins
- Depth Penetration and Detection of pH Gradients in Biofilms by Two-Photon Excitation Microscopy
- Combinatorial discovery of two-photon photoremovable protecting groups