Automated melting curve analysis in droplet microfluidics for single nucleotide polymorphisms (SNP) genotyping
نویسندگان
چکیده
منابع مشابه
Chemical-mediated Melting Curve Analysis for Genotyping of Single Nucleotide Polymorphisms
This report describes a new microfluidic solid-phase chemical-mediated melting curve analysis method for Single Nucleotide Polymorphism (SNP) genotyping. The method is based on allele-specific denaturation of DNA duplexes upon exposure to dynamic chemical gradient. DNA duplexes conjugated on beads are captured in a microfluidic gradient generator device. The device is designed with dams, keepin...
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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...
متن کاملDNA melting analysis for detection of single nucleotide polymorphisms.
BACKGROUND Several methods for detection of single nucleotide polymorphisms (SNPs; e.g., denaturing gradient gel electrophoresis and denaturing HPLC) are indirectly based on the principle of differential melting of heteroduplex DNA. We present a method for detecting SNPs that is directly based on this principle. METHODS We used a double-stranded DNA-specific fluorescent dye, SYBR Green I (SYB...
متن کاملMethods for genotyping single nucleotide polymorphisms.
One of the fruits of the Human Genome Project is the discovery of millions of DNA sequence variants in the human genome. The majority of these variants are single nucleotide polymorphisms (SNPs). A dense set of SNP markers opens up the possibility of studying the genetic basis of complex diseases by population approaches. In all study designs, a large number of individuals must be genotyped wit...
متن کاملMelting curve analysis of SNPs (McSNP): a gel-free and inexpensive approach for SNP genotyping.
High-throughput methods for assaying DNA variation require two important steps: (i) discriminating the variation and (ii) detecting the signal. In this report, we describe a novel SNP genotyping method that we refer to as melting curve analysis of SNPs (McSNP). McSNP combines a classic approach for discriminating alleles, restriction enzyme digestion, with a more recent method for detecting DNA...
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ژورنال
عنوان ژورنال: RSC Advances
سال: 2017
ISSN: 2046-2069
DOI: 10.1039/c6ra26484k