Detection of nucleic acid heteroduplex molecules by denaturing high-performance liquid chromatography and methods for comparative sequencing
The present invention describes a method for separating heteroduplex and homoduplex DNA molecules in a mixture. In the method, such a mixture is applied to a stationary reverse phase support. The heteroduplex and homoduplex molecules are eluted with a mobile phase containing an ion-pairing reagent a...
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| creator | UNDERHILL PETER ANTON OEFNER PETER JOSEF |
| description | The present invention describes a method for separating heteroduplex and homoduplex DNA molecules in a mixture. In the method, such a mixture is applied to a stationary reverse phase support. The heteroduplex and homoduplex molecules are eluted with a mobile phase containing an ion-pairing reagent and an organic solvent. The eluting is carried out under conditions effective to at least partially denature the heteroduplexes (e.g., thermal or chemical denaturing) resulting in the separation of the heteroduplexes from the homoduplexes. The method has many applications including, but not limited to, comparative nucleic acid sequencing, linkage analysis, evolutionary studies, forensics, identification of disease-causing gene mutations, genetic marker development and diagnostics. |
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In the method, such a mixture is applied to a stationary reverse phase support. The heteroduplex and homoduplex molecules are eluted with a mobile phase containing an ion-pairing reagent and an organic solvent. The eluting is carried out under conditions effective to at least partially denature the heteroduplexes (e.g., thermal or chemical denaturing) resulting in the separation of the heteroduplexes from the homoduplexes. The method has many applications including, but not limited to, comparative nucleic acid sequencing, linkage analysis, evolutionary studies, forensics, identification of disease-causing gene mutations, genetic marker development and diagnostics.</abstract><edition>6</edition><oa>free_for_read</oa></addata></record> |
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| subjects | BEER BIOCHEMISTRY CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOIDCHEMISTRY CHEMISTRY COMPOSITIONS OR TEST PAPERS THEREFOR CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES ENZYMOLOGY MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS METALLURGY MICROBIOLOGY MUTATION OR GENETIC ENGINEERING PERFORMING OPERATIONS PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL PROCESSES OF PREPARING SUCH COMPOSITIONS SEPARATION SPIRITS THEIR RELEVANT APPARATUS TRANSPORTING VINEGAR WINE |
| title | Detection of nucleic acid heteroduplex molecules by denaturing high-performance liquid chromatography and methods for comparative sequencing |
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