High-throughput genotyping with infrared fluorescence allele specific hybridization (iFLASH): a simple, reliable and low-cost alternative

Clinical Biochemistry
Thomas M van HimbergenMark Roest

Abstract

To develop and validate a novel genotyping approach, named infrared Fluorescence Allele Specific Hybridization (iFLASH), which combines the principles of allele specific oligonucleotide (ASO) hybridization with the advanced possibilities of infrared imaging. As an example, we genotyped the 55L > M and the 192Q > R common genetic variants of the paraoxonase-1 gene in 92 DNA samples using the iFLASH technique, and validated the outcomes with the restriction fragment length polymorphism (RFLP) and TAQman genotyping assays. There was a 100 percent agreement in genotype outcome among the three methods. Although we found complete unity in genotype outcome, the iFLASH assay has essential advantages over the RFLP and TAQman genotyping assays. First, the iFLASH technique is capable of handling up to 1536 samples per assay, which makes it a suitable technique for high-throughput genotyping. Secondly, because the costs per assay are lower, high-throughput genotyping with iFLASH is affordable.

References

Jan 1, 1983·Proceedings of the National Academy of Sciences of the United States of America·B J ConnerR B Wallace
Jan 1, 1993·Nature Genetics·R HumbertC E Furlong
Mar 20, 1999·Genetic Analysis : Biomolecular Engineering·K J Livak
Jul 15, 2004·Journal of Immunological Methods·Bas B van RijnHieronymus A M Voorbij

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Citations

Dec 22, 2007·Atherosclerosis·Thomas M van HimbergenAnton F H Stalenhoef

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