Rapid hybridization of nucleic acids using isotachophoresis
We use isotachophoresis (ITP) to control and increase the rate of nucleic acid hybridization reactions in free solution. We present a new physical model, validation experiments, and demonstrations of this assay. We studied the coupled physicochemical processes of preconcentration, mixing, and chemic...
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Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 2012-07, Vol.109 (28), p.11127-11132 |
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creator | Bercovici, Moran Han, Crystal M Liao, Joseph C Santiago, Juan G |
description | We use isotachophoresis (ITP) to control and increase the rate of nucleic acid hybridization reactions in free solution. We present a new physical model, validation experiments, and demonstrations of this assay. We studied the coupled physicochemical processes of preconcentration, mixing, and chemical reaction kinetics under ITP. Our experimentally validated model enables a closed form solution for ITP-aided reaction kinetics, and reveals a new characteristic time scale which correctly predicts order 10,000-fold speed-up of chemical reaction rate for order 100 pM reactants, and greater enhancement at lower concentrations. At 500 pM concentration, we measured a reaction time which is 14,000-fold lower than that predicted for standard second-order hybridization. The model and method are generally applicable to acceleration of reactions involving nucleic acids, and may be applicable to a wide range of reactions involving ionic reactants. |
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We present a new physical model, validation experiments, and demonstrations of this assay. We studied the coupled physicochemical processes of preconcentration, mixing, and chemical reaction kinetics under ITP. Our experimentally validated model enables a closed form solution for ITP-aided reaction kinetics, and reveals a new characteristic time scale which correctly predicts order 10,000-fold speed-up of chemical reaction rate for order 100 pM reactants, and greater enhancement at lower concentrations. At 500 pM concentration, we measured a reaction time which is 14,000-fold lower than that predicted for standard second-order hybridization. The model and method are generally applicable to acceleration of reactions involving nucleic acids, and may be applicable to a wide range of reactions involving ionic reactants.</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.1205004109</identifier><identifier>PMID: 22733732</identifier><language>eng</language><publisher>United States: National Academy of Sciences</publisher><subject>Analytical models ; Beacons ; Biological Sciences ; Buffers ; Chemical reactions ; DNA ; DNA - chemistry ; Electrophoresis - methods ; Genetic hybridization ; Humans ; Hybridization ; Isotachophoresis - instrumentation ; Isotachophoresis - methods ; Kinetics ; Liver - metabolism ; mixing ; Modeling ; Models, Chemical ; Models, Statistical ; Normal Distribution ; Nucleic acid hybridization ; Nucleic Acid Hybridization - methods ; Nucleic Acids ; Reactants ; Reaction kinetics ; RNA - chemistry ; Solutions</subject><ispartof>Proceedings of the National Academy of Sciences - PNAS, 2012-07, Vol.109 (28), p.11127-11132</ispartof><rights>copyright © 1993-2008 National Academy of Sciences of the United States of America</rights><rights>Copyright National Academy of Sciences Jul 10, 2012</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c591t-b193fd14203badb66624c42c74c425c64abf8dd629086d42f876219caf4af8733</citedby><cites>FETCH-LOGICAL-c591t-b193fd14203badb66624c42c74c425c64abf8dd629086d42f876219caf4af8733</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://www.pnas.org/content/109/28.cover.gif</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/41684708$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/41684708$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,314,723,776,780,799,881,27903,27904,53769,53771,57995,58228</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/22733732$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Bercovici, Moran</creatorcontrib><creatorcontrib>Han, Crystal M</creatorcontrib><creatorcontrib>Liao, Joseph C</creatorcontrib><creatorcontrib>Santiago, Juan G</creatorcontrib><title>Rapid hybridization of nucleic acids using isotachophoresis</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>We use isotachophoresis (ITP) to control and increase the rate of nucleic acid hybridization reactions in free solution. We present a new physical model, validation experiments, and demonstrations of this assay. We studied the coupled physicochemical processes of preconcentration, mixing, and chemical reaction kinetics under ITP. Our experimentally validated model enables a closed form solution for ITP-aided reaction kinetics, and reveals a new characteristic time scale which correctly predicts order 10,000-fold speed-up of chemical reaction rate for order 100 pM reactants, and greater enhancement at lower concentrations. At 500 pM concentration, we measured a reaction time which is 14,000-fold lower than that predicted for standard second-order hybridization. The model and method are generally applicable to acceleration of reactions involving nucleic acids, and may be applicable to a wide range of reactions involving ionic reactants.</description><subject>Analytical models</subject><subject>Beacons</subject><subject>Biological Sciences</subject><subject>Buffers</subject><subject>Chemical reactions</subject><subject>DNA</subject><subject>DNA - chemistry</subject><subject>Electrophoresis - methods</subject><subject>Genetic hybridization</subject><subject>Humans</subject><subject>Hybridization</subject><subject>Isotachophoresis - instrumentation</subject><subject>Isotachophoresis - methods</subject><subject>Kinetics</subject><subject>Liver - metabolism</subject><subject>mixing</subject><subject>Modeling</subject><subject>Models, Chemical</subject><subject>Models, Statistical</subject><subject>Normal Distribution</subject><subject>Nucleic acid hybridization</subject><subject>Nucleic Acid Hybridization - methods</subject><subject>Nucleic Acids</subject><subject>Reactants</subject><subject>Reaction kinetics</subject><subject>RNA - chemistry</subject><subject>Solutions</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkc1v1DAQxS1ERZeFMycgEhcuaWf8FVuVkFAFFKkSEtCz5TjJrlfZONgJUvnr8bLLFnrqZWak95snex4hLxDOECp2Pg42nSEFAcAR9COyyBVLyTU8JgsAWpWKU35Knqa0AQAtFDwhp5RWjFWMLsjFVzv6pljf1tE3_pedfBiK0BXD7PrWu8I636RiTn5YFT6Fybp1GNchtsmnZ-Sks31qnx_6ktx8_PD98qq8_vLp8-X769IJjVNZo2Zdg5wCq21TSykpd5y6aleFk9zWnWoaSTUo2XDaqUpS1M523OaZsSV5t_cd53rbNq4dpmh7M0a_tfHWBOvN_8rg12YVfhrGtBRMZoO3B4MYfsxtmszWJ9f2vR3aMCeDChhKKegDUKBccgEVZvTNPXQT5jjkS_yhRL67Fpk631MuhpRi2x3fjWB2GZpdhuYuw7zx6t_vHvm_oWWgOAC7zTs7bagyiJi5JXm5RzZpCvHIcJSKV6Cy_nqvdzYYu4o-mZtvFFACIFVaafYbsKO0Ug</recordid><startdate>20120710</startdate><enddate>20120710</enddate><creator>Bercovici, Moran</creator><creator>Han, Crystal M</creator><creator>Liao, Joseph C</creator><creator>Santiago, Juan G</creator><general>National Academy of Sciences</general><general>National Acad Sciences</general><scope>FBQ</scope><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>7QG</scope><scope>7QL</scope><scope>7QP</scope><scope>7QR</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TK</scope><scope>7TM</scope><scope>7TO</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><scope>7S9</scope><scope>L.6</scope><scope>5PM</scope></search><sort><creationdate>20120710</creationdate><title>Rapid hybridization of nucleic acids using isotachophoresis</title><author>Bercovici, Moran ; Han, Crystal M ; Liao, Joseph C ; Santiago, Juan G</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c591t-b193fd14203badb66624c42c74c425c64abf8dd629086d42f876219caf4af8733</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>Analytical models</topic><topic>Beacons</topic><topic>Biological Sciences</topic><topic>Buffers</topic><topic>Chemical reactions</topic><topic>DNA</topic><topic>DNA - chemistry</topic><topic>Electrophoresis - methods</topic><topic>Genetic hybridization</topic><topic>Humans</topic><topic>Hybridization</topic><topic>Isotachophoresis - instrumentation</topic><topic>Isotachophoresis - methods</topic><topic>Kinetics</topic><topic>Liver - metabolism</topic><topic>mixing</topic><topic>Modeling</topic><topic>Models, Chemical</topic><topic>Models, Statistical</topic><topic>Normal Distribution</topic><topic>Nucleic acid hybridization</topic><topic>Nucleic Acid Hybridization - methods</topic><topic>Nucleic Acids</topic><topic>Reactants</topic><topic>Reaction kinetics</topic><topic>RNA - chemistry</topic><topic>Solutions</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Bercovici, Moran</creatorcontrib><creatorcontrib>Han, Crystal M</creatorcontrib><creatorcontrib>Liao, Joseph C</creatorcontrib><creatorcontrib>Santiago, Juan G</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Oncogenes and Growth Factors Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>AGRICOLA</collection><collection>AGRICOLA - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Bercovici, Moran</au><au>Han, Crystal M</au><au>Liao, Joseph C</au><au>Santiago, Juan G</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Rapid hybridization of nucleic acids using isotachophoresis</atitle><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle><addtitle>Proc Natl Acad Sci U S A</addtitle><date>2012-07-10</date><risdate>2012</risdate><volume>109</volume><issue>28</issue><spage>11127</spage><epage>11132</epage><pages>11127-11132</pages><issn>0027-8424</issn><eissn>1091-6490</eissn><abstract>We use isotachophoresis (ITP) to control and increase the rate of nucleic acid hybridization reactions in free solution. We present a new physical model, validation experiments, and demonstrations of this assay. We studied the coupled physicochemical processes of preconcentration, mixing, and chemical reaction kinetics under ITP. Our experimentally validated model enables a closed form solution for ITP-aided reaction kinetics, and reveals a new characteristic time scale which correctly predicts order 10,000-fold speed-up of chemical reaction rate for order 100 pM reactants, and greater enhancement at lower concentrations. At 500 pM concentration, we measured a reaction time which is 14,000-fold lower than that predicted for standard second-order hybridization. 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subjects | Analytical models Beacons Biological Sciences Buffers Chemical reactions DNA DNA - chemistry Electrophoresis - methods Genetic hybridization Humans Hybridization Isotachophoresis - instrumentation Isotachophoresis - methods Kinetics Liver - metabolism mixing Modeling Models, Chemical Models, Statistical Normal Distribution Nucleic acid hybridization Nucleic Acid Hybridization - methods Nucleic Acids Reactants Reaction kinetics RNA - chemistry Solutions |
title | Rapid hybridization of nucleic acids using isotachophoresis |
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