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Amplicon melting analysis with labeled primers: A closed-tube method for differentiating homozygotes and heterozygotes
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Background: Common methods for identification of DNA sequence variants use gel electrophoresis or column separation after PCR.
Methods: We developed a method for sequence variant analysis requiring only PCR and amplicon melting analysis. One of the PCR primers was fluorescently labeled. After PCR, the melting transition of the amplicon was monitored by high-resolution melting analysis. Different homozygotes were distinguished by amplicon melting temperature (T^sub m^). Heterozygotes were identified by low-temperature melting of heteroduplexes, which broadened the overall melting transition. ...
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High-Resolution DNA Melting Analysis for Simultaneous Mutation Scanning and Genotyping in Solution
; ...High-resolution DNA melting analysis with saturation...simultaneous PCR product scanning and probe...of the entire PCR product. For example...for fluorescent melting analysis, both amplification...contrast, fluorescent melting analysis is usually ...
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High-Resolution Melting for Accurate Assessment of DNA Methylation
; ...be resolved by melting analysis in a thermal cycler...T^sup m^ of a PCR product by up to 1 C...high-throughput, in-tube melting analysis. Inclusion in...properties of a PCR product to be examined...methylation by melting analysis relies on the...T^sub m^ ...
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Sensitivity and Specificity of Single-Nucleotide Polymorphism Scanning by High-Resolution Melting Analysis
; ...High-resolution melting analysis with the dye LCGreen...depends on the PCR product size and the type...variant within a PCR product would provide...high-resolution melting analysis. The technique...highresolution melting analysis for mutation ...
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Detection of Factor VIII Gene Mutations by High-Resolution Melting Analysis
; ...Methods: We evaluated high-resolution melting analysis as an option for screening for F8...normal amplicons to assess whether melting analysis could detect these variants. We examined...variants we examined were resolved by melting analysis. For the other 2 mutations, the melting...
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Rapid and Sensitive Detection of BRCA1/2 Mutations in a Diagnostic Setting: Comparison of Two High-Resolution Melting Platforms
; ...High-resolution melting is an emerging technique...accurate high-resolution melting analysis possible and created...High-resolution melting analysis was performed on...High-resolution melting analysis of nucleic acids...amplicons in the PCR product, vs only ...
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Two for Typing: Homogeneous Combined Single-Nucleotide Polymorphism Scanning and Genotyping
; ...probe-based PCR product identification...melting of a whole PCR product and an unlabeled...melt the whole PCR product without probes...transitions of a whole PCR product from a wild...genotyping solely by melting analysis was demonstrated...of ...
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Fluorescent SNP Genotyping by High-Resolution Melting Analysis without Probes
; 12 Fluorescent SNP Genotyping by High-Resolution Melting Analysis without Probes. Michael Liew, Mark Johnson, Robert...were developed using high-resolution fluorescent melting analysis of small amplicons. Small amplicons were rapidly...
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High-resolution genotyping by amplicon melting analysis using LCGreen
; Background: High-resolution amplicon melting analysis was recently introduced as a closed-tu...conditions. Conclusions: High-resolution melting analysis of PCR products amplified in the presence...SYBR Green I was used for amplicon melting analysis and provided a rough ...
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Quadruplex Genotyping of F5, F2, and MTHFR Variants in a Single Closed Tube by High-Resolution Amplicon Melting
; ...analysis requires high-resolution melting analysis and controlled reaction conditions...and then conducted a high-resolution melting analysis. Heterozygotes were identified at...Rapid-cycle PCR with high-resolution melting analysis allows simple and accurate multiplex...
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Instrument Comparison for Heterozygote Scanning of Single and Double Heterozygotes: A Correction and Extension of Herrmann et al., Clin Chem 2006;52:494-503
; ...further shows the capability of DNA melting analysis and its dependency on instrument resolution...unique sequence variants. Aspects of melting analysis are covered by issued and pending...Wittwer CT. Voelkerding KV. Amplicon DNA melting analysis for mutation scanning and ...
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