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Chemical gradient‐mediated melting curve analysis for genotyping of SNPs
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| Zeitschriftentitel: | ELECTROPHORESIS |
|---|---|
| Personen und Körperschaften: | , , |
| In: | ELECTROPHORESIS, 30, 2009, 14, S. 2536-2543 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
Wiley
|
| Schlagwörter: |
| author_facet |
Russom, Aman Irimia, Daniel Toner, Mehmet Russom, Aman Irimia, Daniel Toner, Mehmet |
|---|---|
| author |
Russom, Aman Irimia, Daniel Toner, Mehmet |
| spellingShingle |
Russom, Aman Irimia, Daniel Toner, Mehmet ELECTROPHORESIS Chemical gradient‐mediated melting curve analysis for genotyping of SNPs Clinical Biochemistry Biochemistry Analytical Chemistry |
| author_sort |
russom, aman |
| spelling |
Russom, Aman Irimia, Daniel Toner, Mehmet 0173-0835 1522-2683 Wiley Clinical Biochemistry Biochemistry Analytical Chemistry http://dx.doi.org/10.1002/elps.200800729 <jats:title>Abstract</jats:title><jats:p>This report describes a microfluidic solid‐phase chemical gradient‐mediated melting curve analysis method for SNP analysis. The method is based on allele‐specific denaturation to discriminate mismatched (MM) from perfectly matched (PM) DNA duplexes upon exposure to linear chemical gradient. PM and MM DNA duplexes conjugated on beads are captured in a microfluidic gradient generator device designed with dams, keeping the beads trapped perpendicular to a gradient generating channel. Two denaturants, formamide and urea, were tested for their ability to destabilize the DNA duplex by competing with Watson–Crick pairing. Upon exposure to the chemical gradient, rapid denaturing profile was monitored in real time using fluorescence microscopy. The results show that the two duplexes exhibit different kinetics of denaturation profiles, enabling discrimination of MM from PM DNA duplexes to score SNP.</jats:p> Chemical gradient‐mediated melting curve analysis for genotyping of SNPs ELECTROPHORESIS |
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10.1002/elps.200800729 |
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| title |
Chemical gradient‐mediated melting curve analysis for genotyping of SNPs |
| title_unstemmed |
Chemical gradient‐mediated melting curve analysis for genotyping of SNPs |
| title_full |
Chemical gradient‐mediated melting curve analysis for genotyping of SNPs |
| title_fullStr |
Chemical gradient‐mediated melting curve analysis for genotyping of SNPs |
| title_full_unstemmed |
Chemical gradient‐mediated melting curve analysis for genotyping of SNPs |
| title_short |
Chemical gradient‐mediated melting curve analysis for genotyping of SNPs |
| title_sort |
chemical gradient‐mediated melting curve analysis for genotyping of snps |
| topic |
Clinical Biochemistry Biochemistry Analytical Chemistry |
| url |
http://dx.doi.org/10.1002/elps.200800729 |
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2009 |
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2536-2543 |
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<jats:title>Abstract</jats:title><jats:p>This report describes a microfluidic solid‐phase chemical gradient‐mediated melting curve analysis method for SNP analysis. The method is based on allele‐specific denaturation to discriminate mismatched (MM) from perfectly matched (PM) DNA duplexes upon exposure to linear chemical gradient. PM and MM DNA duplexes conjugated on beads are captured in a microfluidic gradient generator device designed with dams, keeping the beads trapped perpendicular to a gradient generating channel. Two denaturants, formamide and urea, were tested for their ability to destabilize the DNA duplex by competing with Watson–Crick pairing. Upon exposure to the chemical gradient, rapid denaturing profile was monitored in real time using fluorescence microscopy. The results show that the two duplexes exhibit different kinetics of denaturation profiles, enabling discrimination of MM from PM DNA duplexes to score SNP.</jats:p> |
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| author | Russom, Aman, Irimia, Daniel, Toner, Mehmet |
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| description | <jats:title>Abstract</jats:title><jats:p>This report describes a microfluidic solid‐phase chemical gradient‐mediated melting curve analysis method for SNP analysis. The method is based on allele‐specific denaturation to discriminate mismatched (MM) from perfectly matched (PM) DNA duplexes upon exposure to linear chemical gradient. PM and MM DNA duplexes conjugated on beads are captured in a microfluidic gradient generator device designed with dams, keeping the beads trapped perpendicular to a gradient generating channel. Two denaturants, formamide and urea, were tested for their ability to destabilize the DNA duplex by competing with Watson–Crick pairing. Upon exposure to the chemical gradient, rapid denaturing profile was monitored in real time using fluorescence microscopy. The results show that the two duplexes exhibit different kinetics of denaturation profiles, enabling discrimination of MM from PM DNA duplexes to score SNP.</jats:p> |
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| spelling | Russom, Aman Irimia, Daniel Toner, Mehmet 0173-0835 1522-2683 Wiley Clinical Biochemistry Biochemistry Analytical Chemistry http://dx.doi.org/10.1002/elps.200800729 <jats:title>Abstract</jats:title><jats:p>This report describes a microfluidic solid‐phase chemical gradient‐mediated melting curve analysis method for SNP analysis. The method is based on allele‐specific denaturation to discriminate mismatched (MM) from perfectly matched (PM) DNA duplexes upon exposure to linear chemical gradient. PM and MM DNA duplexes conjugated on beads are captured in a microfluidic gradient generator device designed with dams, keeping the beads trapped perpendicular to a gradient generating channel. Two denaturants, formamide and urea, were tested for their ability to destabilize the DNA duplex by competing with Watson–Crick pairing. Upon exposure to the chemical gradient, rapid denaturing profile was monitored in real time using fluorescence microscopy. The results show that the two duplexes exhibit different kinetics of denaturation profiles, enabling discrimination of MM from PM DNA duplexes to score SNP.</jats:p> Chemical gradient‐mediated melting curve analysis for genotyping of SNPs ELECTROPHORESIS |
| spellingShingle | Russom, Aman, Irimia, Daniel, Toner, Mehmet, ELECTROPHORESIS, Chemical gradient‐mediated melting curve analysis for genotyping of SNPs, Clinical Biochemistry, Biochemistry, Analytical Chemistry |
| title | Chemical gradient‐mediated melting curve analysis for genotyping of SNPs |
| title_full | Chemical gradient‐mediated melting curve analysis for genotyping of SNPs |
| title_fullStr | Chemical gradient‐mediated melting curve analysis for genotyping of SNPs |
| title_full_unstemmed | Chemical gradient‐mediated melting curve analysis for genotyping of SNPs |
| title_short | Chemical gradient‐mediated melting curve analysis for genotyping of SNPs |
| title_sort | chemical gradient‐mediated melting curve analysis for genotyping of snps |
| title_unstemmed | Chemical gradient‐mediated melting curve analysis for genotyping of SNPs |
| topic | Clinical Biochemistry, Biochemistry, Analytical Chemistry |
| url | http://dx.doi.org/10.1002/elps.200800729 |