scholarly journals Sub-7-second genotyping of single-nucleotide polymorphism by high-resolution melting curve analysis on a thermal digital microfluidic device

Lab on a Chip ◽  
2016 ◽  
Vol 16 (4) ◽  
pp. 743-752 ◽  
Author(s):  
Tianlan Chen ◽  
Yanwei Jia ◽  
Cheng Dong ◽  
Jie Gao ◽  
Pui-In Mak ◽  
...  
Keyword(s):  
High Resolution Melting ◽  
Melting Curve ◽  
Curve Analysis ◽  
Melting Curve Analysis ◽  
Dna Melting ◽  
Nucleotide Polymorphism ◽  
Single Nucleotide ◽  
High Resolution Melting Curve ◽  
Digital Microfluidic ◽  
Multiple Samples

A novel thermal digital microfluidic (T-DMF) device enables precise thermal modulation and pipelined measurement of multiple samples. Ultrafast DNA melting curve analysis is achieved in less than 7 seconds, with the resolution adequate for single-nucleotide discrimination.

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.

One-shot high-resolution melting curve analysis for KRAS point-mutation discrimination on a digital microfluidics platform

Lab on a Chip ◽  
2022 ◽  
Author(s):  
Mingzhong Li ◽  
Liang Wan ◽  
Man-Kay Law ◽  
Li Meng ◽  
Yanwei Jia ◽  
...  
Keyword(s):  
High Resolution ◽  
High Resolution Melting ◽  
Digital Microfluidics ◽  
Melting Curve ◽  
High Accuracy ◽  
Curve Analysis ◽  
Melting Curve Analysis ◽  
Single Nucleotide ◽  
High Resolution Melting Curve ◽  
Microfluidics Platform

Proposed high-accuracy one-shot melting curve analysis on a digital microfluidics platform, enabling KRAS gene discrimination with single-nucleotide mutations in 3 seconds.

scholarly journals Interleukin-7 receptor Thr244Ile gene polymorphism and the risk of systemic lupus erythematosus

2016 ◽  
Vol 85 (3) ◽  
pp. 192 ◽  
Author(s):  
Pawel Piotr Jagodzinski ◽  
Piotr Piotrowski ◽  
Marzena Olesińska
Keyword(s):  
Lupus Erythematosus ◽  
High Resolution Melting ◽  
Melting Curve ◽  
Melting Curve Analysis ◽  
Neurological Manifestations ◽  
Nucleotide Polymorphism ◽  
Single Nucleotide ◽  
Interleukin 7 ◽  
High Resolution Melting Curve ◽  
And Control

Aim. Recently, the IL-7 receptor (IL-7R) C>T (rs6897932) single nucleotide polymorphism (SNP), which causes a Thr244Ile substitution in the IL-7R α‑chain, has been suggested as a risk factor for SLE.Material and Methods. Using high‑resolution melting curve analysis we studied the distribution of the IL-7R C>T polymorphism in SLE patients (n = 281) and control subjects (n = 541) in the Polish population.Results. We did not find significant differences in the distribution of the IL-7R C>T genotype and alleles between SLE patients and controls. However, in the dominant model (T/T and C/T vs C/C genotypes), we observed a protective effect of the IL-7R C>T polymorphism against the presence of neurological manifestations of SLE [OR = 0.3631 (95% CI = 0.1895–0.6954), p = 0.0017, pcorr = 0.0323] and the presence of anti‑Scl-70 antibodies (Ab) [OR = 0.3141 (95% CI = 0.1503–0.6561), p = 0.0014, pcorr = 0.0266].Conclusion. Our studies suggest that the IL-7R C>T (rs6897932) polymorphism might be involved in the neurological manifestations and the presence of anti‑Scl-70 Abs in patients with SLE.

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.

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