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• W67642703 abstract "Fluorescent nucleic acid detection in polymerase chain reaction (PCR) generally uses oligonucleotide probes labeled with covalently attached dyes. However, unlabeled oligonucleotides in the presence of saturating DNA dyes can also serve as hybridization probes. The DNA dye, LCGreen Plus, and a 3'-blocked unlabeled probe are added before amplification, and asymmetric PCR is performed at a 1:5 to 1:10 primer ratio. After PCR is complete, fluorescent melting curves reveal both probe melting at low temperature and amplicon melting at high temperature. After background removal, the melting temperature(s) of the probe/target duplex specific to the allele(s) amplified are revealed. Probes between 20 and 40 bp with T(m)s between 50 and 85 degrees C are effective. The method requires only three standard oligonucleotides and endpoint fluorescence melting. No real-time PCR or allele-specific amplification is needed. Unlabeled probes are inexpensive, provide the sequence specificity of probes, and allow simultaneous identification of multiple alleles by melting analysis." @default.
• W67642703 created "2016-06-24" @default.
• W67642703 creator A5018572080 @default.
• W67642703 creator A5031374595 @default.
• W67642703 creator A5051507868 @default.
• W67642703 date "2008-01-01" @default.
• W67642703 modified "2023-10-02" @default.
• W67642703 title "SNP Genotyping by Unlabeled Probe Melting Analysis" @default.
• W67642703 cites W1985356937 @default.
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• W67642703 doi "https://doi.org/10.1007/978-1-60327-040-3_14" @default.
• W67642703 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/18695968" @default.
• W67642703 hasPublicationYear "2008" @default.
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