Abstract
Real-time polymerase chain reaction (PCR) has become a standard tool in both quantitative gene expression and genetic variation analysis. Data collection is performed throughout the PCR process, thus combining amplification and detection into a single step. This can be achieved by combining a variety of different fluorescent chemistries that correlate the concentration of an amplified PCR product to changes in fluorescence intensity. Hybridization probe pairs and single-labeled probes are sequence-specific, dye-labeled oligonucleotides, used in real-time PCR approaches, in particular for genotyping of single nucleotide polymorphisms (SNPs). In that case, a detector probe is designed to cover the polymorphism. Allelic variants are identified and differentiated via post-PCR melting curve analysis. A single melting curve can distinguish different T ms, and differently labeled probes may be used, theoretically allowing multiplexed genotyping of several SNPs.
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http://www.ncbi.nlm.nih.gov. SNP database accession number: rs1799964.
http://www.ncbi.nlm.nih.gov. SNP database accession numbers: rs1799724, rs1800630rs.
http://www.ncbi.nlm.nih.gov. Gene bank accession numbers: gi 26449126 gb AC096632.3 (ADIPOR1; SNP at position 55649), gi 4165008 gb AC005343.1 (ADIPOR2; SNP at position 91846), gi 4165008 gb AC005343.1 (ADIPOR2; SNP at position 89539).
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© 2008 Humana Press Inc., Totowa, NJ
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Froehlich, T., Geulen, O. (2008). Hybridization Probe Pairs and Single-Labeled Probes: an Alternative Approach for Genotyping and Quantification. In: Marx, A., Seitz, O. (eds) Molecular Beacons: Signalling Nucleic Acid Probes, Methods, and Protocols. Methods in Molecular Biology, vol 429. Humana Press. https://doi.org/10.1007/978-1-60327-040-3_9
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DOI: https://doi.org/10.1007/978-1-60327-040-3_9
Publisher Name: Humana Press
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