ArticleNucleic acids research2004
A new approach to SNP genotyping with fluorescently labeled mononucleotides.
Article in Nucleic acids research, 2004. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
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Who cites it
6 citing papers in PubMed, 32 citations in OpenAlex.
- The development of peptide-boron difluoride formazanate conjugates as fluorescence imaging agents.RSC advances · 2020Article
- Rice molecular breeding laboratories in the genomics era: Current status and future considerations.International journal of plant genomics · 2008Article
- Reliable allele detection using SNP-based PCR primers containing Locked Nucleic Acid: application in genetic mapping.Plant methods · 2007Article
- Homogeneous point mutation detection by quantum dot-mediated two-color fluorescence coincidence analysis.Nucleic acids research · 2006Article
- Flow cytometric detection of specific RNAs in native human cells with quenched autoligating FRET probes.Proceedings of the National Academy of Sciences of the United States of America · 2006Article
- A FRET-based analysis of SNPs without fluorescent probes.Nucleic acids research · 2004Article
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Fluorescence resonance energy transfer (FRET) is one of the most powerful and promising tools for single nucleotide polymorphism (SNP) genotyping. However, the present methods using FRET require expensive reagents such as fluorescently labeled oligonucleotides. Here, we describe a novel and cost-effective method for SNP genotyping using FRET. The technique is based on allele-specific primer extension using mononucleotides labeled with a green dye and a red dye. When the target DNA contains the sequence complementary to the primer, extension of the primer incorporates the green and red dye-labeled nucleotides into the strand, and red fluorescence is emitted by FRET. In contrast, when the 3' end nucleotide of the primer is not complementary to the target DNA, there is no extension of the primer, or FRET signal. Therefore, discrimination among genotypes is achieved by measuring the intensity of red fluorescence after the extension reaction. We have validated this method with 11 SNPs, which were successfully determined by end-point measurements of fluorescence intensity. The new strategy is simple and cost-effective, because all steps of the preparation consist of simple additions of solutions and incubation, and the dye-labeled mononucleotides are applicable to all SNP analyses. This method will be suitable for large-scale genotyping.
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