HRM for High Throughput Applications
Tuesday, November 19, 2013
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Chapters
Introduction
What is HRM ?
Amplicon Melt Profile Differences are Key
Developments by Roche
Critical Dye Chemistry Progress
Roche Dye
Single Nucleotide Polymorphisms
Detection of Carriers
What is a SNP ?
Theoretical Profiles
In Practice
What amplicons comprise a heterozygous peak ?
Designing an HRM Assay for a High Throughput Setting
Data differences must be greater than inter-sample variations
Assay Optimization
Parameters to Control
Amplicon Melting
Selection of Primers
Synthetic Amplification Template
Primer and Template Modifications
Basic PCR Optimization
Example Assays
96 Samples in Triplicate
Unique Outcomes
Tips, Tools, and Tricks
SNP Specific Limitations
Software Specific Limitations
Comparison of Three Analysis Methods
Conclusions
Acknowledgements
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