NDLI: Automated melting curve analysis in droplet microfluidics for single nucleotide polymorphisms (SNP) genotyping

Content Provider	Royal Society of Chemistry (RSC)
Author	Lu, Y.-W. Jang, J.-S. R. Liu, F.-W. Lin, E.-C. Ding, S.-T.
Copyright Year	2017
Abstract	An integrated microchip platform with automated analysis capability for DNA melting curves is developed for Single Nucleotide Polymorphism (SNP) genotyping applications. The microchip contains fluidic channels where genomic DNA samples are encapsulated into a series of droplets and transported through a detection region with a stable temperature gradient. As the temperature is elevated from 60 °C to 85 °C, the DNA strains denature and the associated fluorescence signals decay with this relationship being acquired as the melting curve. The droplets serve as discrete reactors to conduct DNA melting curve analysis in the liquid phase, eliminating the need for immobilization of reagents. They provide the advantage of signal homogeneity, which reduces the signal fluctuations, and thus the signal-to-noise ratio is improved. In addition, a droplet detection and tracking software system which can identify the droplets, records the fluorescence intensities, plots the melting curves, and finds the melting temperatures, is developed making automated SNP genotyping possible. The platform has been verified with genomic DNA from Landrace sows and shown successful SNP discrimination from homozygotes and heterozygotes which demonstrates its potential to conduct on-site SNP genotyping for disease research, medical diagnostics, agriculture, and farm animal reproduction.
Starting Page	4646
Ending Page	4655
Page Count	10
File Format	HTM / HTML PDF
ISSN	20462069
Volume Number	7
Issue Number	8
Journal	RSC Advances
DOI	10.1039/c6ra26484k
Language	English
Publisher	Royal Society of Chemistry
Access Restriction	Open
Subject Keyword	SNP Genotyping DNA Integrated circuit Fluorescence Signal-to-noise ratio Melting curve analysis Single-nucleotide polymorphism Landrace SNP genotyping Nucleic acid thermodynamics
Content Type	Text
Resource Type	Article
Subject	Chemistry Chemical Engineering

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