Thermoelectric method for sequencing DNA
This study describes a novel, thermoelectric method for DNA sequencing in a microfluidic device. The method measures the heat released when DNA polymerase inserts a deoxyribonucleoside triphosphate into a primed DNA template. The study describes the principle of operation of a laminar flow microflui...
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| Veröffentlicht in: | Lab on a chip 2011-05, Vol.11 (10), p.1761-1769 |
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| container_end_page | 1769 |
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| container_title | Lab on a chip |
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| creator | Nestorova, Gergana G Guilbeau, Eric J |
| description | This study describes a novel, thermoelectric method for DNA sequencing in a microfluidic device. The method measures the heat released when DNA polymerase inserts a deoxyribonucleoside triphosphate into a primed DNA template. The study describes the principle of operation of a laminar flow microfluidic chip with a reaction zone that contains DNA template/primer complex immobilized to the inner surface of the device's lower channel wall. A thin-film thermopile attached to the external surface of the lower channel wall measures the dynamic change in temperature that results when Klenow polymerase inserts a deoxyribonucleoside triphosphate into the DNA template. The intrinsic rejection of common-mode thermal signals by the thermopile in combination with hydrodynamic focused flow allows for the measurement of temperature changes on the order of 10(-4) K without control of ambient temperature. To demonstrate the method, we report the sequencing of a model oligonucleotide containing 12 bases. Results demonstrate that it is feasible to sequence DNA by measuring the heat released during nucleotide incorporation. This thermoelectric method for sequencing DNA may offer a novel new method of DNA sequencing for personalized medicine applications. |
| doi_str_mv | 10.1039/c0lc00733a |
| format | Article |
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The method measures the heat released when DNA polymerase inserts a deoxyribonucleoside triphosphate into a primed DNA template. The study describes the principle of operation of a laminar flow microfluidic chip with a reaction zone that contains DNA template/primer complex immobilized to the inner surface of the device's lower channel wall. A thin-film thermopile attached to the external surface of the lower channel wall measures the dynamic change in temperature that results when Klenow polymerase inserts a deoxyribonucleoside triphosphate into the DNA template. The intrinsic rejection of common-mode thermal signals by the thermopile in combination with hydrodynamic focused flow allows for the measurement of temperature changes on the order of 10(-4) K without control of ambient temperature. To demonstrate the method, we report the sequencing of a model oligonucleotide containing 12 bases. Results demonstrate that it is feasible to sequence DNA by measuring the heat released during nucleotide incorporation. 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The method measures the heat released when DNA polymerase inserts a deoxyribonucleoside triphosphate into a primed DNA template. The study describes the principle of operation of a laminar flow microfluidic chip with a reaction zone that contains DNA template/primer complex immobilized to the inner surface of the device's lower channel wall. A thin-film thermopile attached to the external surface of the lower channel wall measures the dynamic change in temperature that results when Klenow polymerase inserts a deoxyribonucleoside triphosphate into the DNA template. The intrinsic rejection of common-mode thermal signals by the thermopile in combination with hydrodynamic focused flow allows for the measurement of temperature changes on the order of 10(-4) K without control of ambient temperature. To demonstrate the method, we report the sequencing of a model oligonucleotide containing 12 bases. Results demonstrate that it is feasible to sequence DNA by measuring the heat released during nucleotide incorporation. This thermoelectric method for sequencing DNA may offer a novel new method of DNA sequencing for personalized medicine applications.</description><subject>Conserved sequence</subject><subject>DNA - chemistry</subject><subject>DNA Primers - chemistry</subject><subject>DNA-Directed DNA Polymerase - metabolism</subject><subject>Microfluidic Analytical Techniques - instrumentation</subject><subject>Microfluidic Analytical Techniques - methods</subject><subject>Sequence Analysis, DNA - instrumentation</subject><subject>Sequence Analysis, DNA - methods</subject><subject>Temperature</subject><issn>1473-0197</issn><issn>1473-0189</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkD1PwzAURS0EoqWw8ANQNhBSwM_Pqe2xKp9SBUuZo8R-pkFJU-xk4N8T1FJGpvuGo3OfLmPnwG-Ao7m1vLacK8TigI1BKkw5aHO4v40asZMYPziHTE71MRsJGBINH7Or5YpC01JNtguVTRrqVq1LfBuSSJ89rW21fk_uXman7MgXdaSzXU7Y28P9cv6ULl4fn-ezRWoRdJcaAkSXFaRAe-HAOWGlmypdkkc9PFl44DrTEhBU5qWwSktdghZFSUiEE3a59W5CO_THLm-qaKmuizW1fcwNV6BwEP5L6qkEIwQXA3m9JW1oYwzk802omiJ85cDznwnzvwkH-GKn7cuG3B793Qy_AV1jagQ</recordid><startdate>20110521</startdate><enddate>20110521</enddate><creator>Nestorova, Gergana G</creator><creator>Guilbeau, Eric J</creator><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>7TM</scope></search><sort><creationdate>20110521</creationdate><title>Thermoelectric method for sequencing DNA</title><author>Nestorova, Gergana G ; Guilbeau, Eric J</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c318t-9e133d5ae718f2d1dd2c4d678bef38073af10858413175f42c7848b182abe3ee3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>Conserved sequence</topic><topic>DNA - chemistry</topic><topic>DNA Primers - chemistry</topic><topic>DNA-Directed DNA Polymerase - metabolism</topic><topic>Microfluidic Analytical Techniques - instrumentation</topic><topic>Microfluidic Analytical Techniques - methods</topic><topic>Sequence Analysis, DNA - instrumentation</topic><topic>Sequence Analysis, DNA - methods</topic><topic>Temperature</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Nestorova, Gergana G</creatorcontrib><creatorcontrib>Guilbeau, Eric J</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Nucleic Acids Abstracts</collection><jtitle>Lab on a chip</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Nestorova, Gergana G</au><au>Guilbeau, Eric J</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Thermoelectric method for sequencing DNA</atitle><jtitle>Lab on a chip</jtitle><addtitle>Lab Chip</addtitle><date>2011-05-21</date><risdate>2011</risdate><volume>11</volume><issue>10</issue><spage>1761</spage><epage>1769</epage><pages>1761-1769</pages><issn>1473-0197</issn><eissn>1473-0189</eissn><abstract>This study describes a novel, thermoelectric method for DNA sequencing in a microfluidic device. 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| ispartof | Lab on a chip, 2011-05, Vol.11 (10), p.1761-1769 |
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| language | eng |
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| source | MEDLINE; Royal Society Of Chemistry Journals 2008-; Alma/SFX Local Collection |
| subjects | Conserved sequence DNA - chemistry DNA Primers - chemistry DNA-Directed DNA Polymerase - metabolism Microfluidic Analytical Techniques - instrumentation Microfluidic Analytical Techniques - methods Sequence Analysis, DNA - instrumentation Sequence Analysis, DNA - methods Temperature |
| title | Thermoelectric method for sequencing DNA |
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