DNA methylation assay using droplet-based DNA melting curve analysis

Lab on a Chip ◽  
2018 ◽  
Vol 18 (3) ◽  
pp. 514-521 ◽  
Author(s):  
F.-W. Liu ◽  
H.-F. Liao ◽  
S.-P. Lin ◽  
Y.-W. Lu
Keyword(s):  
Dna Methylation ◽  
Methylation Status ◽  
Melting Curve ◽  
Genomic Region ◽  
Curve Analysis ◽  
Droplet Microfluidics ◽  
Melting Curve Analysis ◽  
Dna Melting ◽  
Methylation Assay ◽  
Specific Genomic Region

A novel platform, combining droplet microfluidics and melting curve analysis, was developed to detect and to quantify the methylation status in a specific genomic region.

scholarly journals Automated melting curve analysis in droplet microfluidics for single nucleotide polymorphisms (SNP) genotyping

RSC Advances ◽  
2017 ◽  
Vol 7 (8) ◽  
pp. 4646-4655 ◽  
Author(s):  
F.-W. Liu ◽  
S.-T. Ding ◽  
E.-C. Lin ◽  
Y.-W. Lu ◽  
J.-S. R. Jang
Keyword(s):  
Melting Curve ◽  
Automated Analysis ◽  
Curve Analysis ◽  
Snp Genotyping ◽  
Droplet Microfluidics ◽  
Melting Curve Analysis ◽  
Dna Melting ◽  
Nucleotide Polymorphisms ◽  
Nucleotide Polymorphism ◽  
Single Nucleotide

An integrated microchip platform with automated analysis capability for DNA melting curves is developed for Single Nucleotide Polymorphism (SNP) genotyping applications.

Heat transfer time determination based on DNA melting curve analysis

2019 ◽  
Vol 24 (1) ◽  
Author(s):  
Hanliang Zhu ◽  
Huanan Li ◽  
Haoqing Zhang ◽  
Zdenka Fohlerova ◽  
Sheng Ni ◽  
...  
Keyword(s):  
Heat Transfer ◽  
Melting Curve ◽  
Curve Analysis ◽  
Melting Curve Analysis ◽  
Dna Melting ◽  
Transfer Time ◽  
Time Determination

High-resolution DNA melting curve analysis to establish HLA genotypic identity

2004 ◽  
Vol 64 (2) ◽  
pp. 156-164 ◽  
Author(s):  
L. Zhou ◽  
J. Vandersteen ◽  
L. Wang ◽  
T. Fuller ◽  
M. Taylor ◽  
...  
Keyword(s):  
High Resolution ◽  
Melting Curve ◽  
Curve Analysis ◽  
Melting Curve Analysis ◽  
Dna Melting

scholarly journals High-Resolution Melting Curve Analysis of Genomic and Whole-Genome Amplified DNA

2008 ◽  
Vol 54 (12) ◽  
pp. 2055-2058 ◽  
Author(s):  
Michael H Cho ◽  
Dawn Ciulla ◽  
Barbara J Klanderman ◽  
Benjamin A Raby ◽  
Edwin K Silverman
Keyword(s):  
High Resolution ◽  
High Resolution Melting ◽  
Melting Curve ◽  
Curve Analysis ◽  
Melting Curve Analysis ◽  
Dna Melting ◽  
Whole Genome ◽  
Nucleotide Polymorphisms ◽  
Wild Type ◽  
High Resolution Melting Curve

Abstract Background: High-resolution melting curve analysis is an accurate method for mutation detection in genomic DNA. Few studies have compared the performance of high-resolution DNA melting curve analysis (HRM) in genomic and whole-genome amplified (WGA) DNA. Methods: In 39 paired genomic and WGA samples, 23 amplicons from 9 genes were PCR amplified and analyzed by high-resolution melting curve analysis using the 96-well LightScanner (Idaho Technology). We used genotyping and bidirectional resequencing to verify melting curve results. Results: Melting patterns were concordant between the genomic and WGA samples in 823 of 863 (95%) analyzed sample pairs. Of the discordant patterns, there was an overrepresentation of alternate melting curve patterns in the WGA samples, suggesting the presence of a mutation (false positives). Targeted resequencing in 135 genomic and 136 WGA samples revealed 43 single nucleotide polymorphisms (SNPs). All SNPs detected in genomic samples were also detected in WGA. Additional genotyping and sequencing allowed the classification of 628 genomic and 614 WGA amplicon samples. Heterozygous variants were identified by non–wild-type melting pattern in 98% of genomic and 97% of WGA samples (P = 0.11). Wild types were correctly classified in 99% of genomic and 91% of WGA samples (P < 0.001). Conclusions: In WGA DNA, high-resolution DNA melting curve analysis is a sensitive tool for SNP discovery through detection of heterozygote variants; however, it may misclassify a greater number of wild-type samples.

scholarly journals Rapid, Simple, and Accurate Detection of K-ras Mutations From Body Fluids Using Real-Time PCR and DNA Melting Curve Analysis

2006 ◽  
Vol 37 (5) ◽  
pp. 286-289 ◽  
Author(s):  
Sayaka Mori ◽  
Kazuyuki Sugahara ◽  
Akiko Uemura ◽  
Norihiko Akamatsu ◽  
Ryuzi Tutsumi ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Melting Curve ◽  
Body Fluids ◽  
Curve Analysis ◽  
Melting Curve Analysis ◽  
Dna Melting ◽  
Ras Mutations ◽  
Accurate Detection

Promoter Methylation and Gene Expression of Pin1 Associated with the Risk of Alzheimer’s Disease in Southern Chinese

2021 ◽  
Vol 17 (13) ◽  
pp. 1232-1237
Author(s):  
Suk L. Ma ◽  
Nelson L.S. Tang ◽  
Linda C. Wa Lam
Keyword(s):  
Gene Expression ◽  
Alzheimer’S Disease ◽  
Alzheimer's Disease ◽  
Dna Methylation ◽  
Protective Factor ◽  
Melting Curve ◽  
Melting Curve Analysis ◽  
Southern Chinese ◽  
Methylation Assay ◽  
Normal Controls

Background: Pin1 is a propyl cis-trans isomerase and it has been associated with age-atonset of Alzheimer’s disease (AD) and other pathological characteristics of AD. DNA methylation is one of the gene regulation mechanisms and it might affect the gene expression. Objectives: This study was aimed to examine the correlation between DNA methylation and gene expression of Pin1 and its effect on the risk of AD in a Chinese population. Methods: 80 AD patients and 180 normal controls were recruited in this study and their cognitive functions were assessed. Pin1 gene expression and methylation were quantified by real-time RT-PCR and Melting Curve Analysis-Methylation assay (MCA-Meth), respectively. Results: Our finding revealed a positive correlation between methylation and gene expression of Pin1 (p=0.001) and increased Pin1 methylation was predisposed to the risk of AD (p<0.001). CG genotype of Pin1 SNP rs2287839 was associated with higher gene expression of Pin1 (p=0.036) and the effect was only prominent in normal controls as AD patients were already methylated at Pin1 promoter. Furthermore, methylation of Pin1 was associated with better performance in cognition (p=0.018). Conclusion: Our result further supported the involvement of Pin1 in AD and the increased level of Pin1 might be a protective factor for AD.

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