Highly sensitive amperometric detection of genomic DNA in animal tissues
A simple and highly sensitive method for the detection of genomic DNA in tissue samples is described. It is based on amperometric detection of target DNA by forming an analyte/polymeric activator bilayer on a gold electrode. The biotinylated target DNA is hybridized to oligonucleotide capture probes...
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| Veröffentlicht in: | Nucleic acids research 2004-01, Vol.32 (2), p.e15-e15 |
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| creator | Xie, Hong Yu, Yuan Hong Mao, Pei‐Lin Gao, Zhiqiang |
| description | A simple and highly sensitive method for the detection of genomic DNA in tissue samples is described. It is based on amperometric detection of target DNA by forming an analyte/polymeric activator bilayer on a gold electrode. The biotinylated target DNA is hybridized to oligonucleotide capture probes immobilized on the gold electrode, forming the first layer. A subsequent binding of glucose oxidase– avidin conjugate to the target DNA and the introduction of a second layer of a redox polymer to the electrode, via layer‐by‐layer electrostatic self‐assembly, allow for electrochemical detection of the catalytic oxidation current of glucose in a PBS solution. Less than 2.0 fg of rat genomic DNA, for both regulated and house‐keeping genes, can be easily detected in 2.5 µl droplets. The proposed procedure shows very high specificity for genomic DNA in a RT–PCR mixture. |
| doi_str_mv | 10.1093/nar/gnh016 |
| format | Article |
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It is based on amperometric detection of target DNA by forming an analyte/polymeric activator bilayer on a gold electrode. The biotinylated target DNA is hybridized to oligonucleotide capture probes immobilized on the gold electrode, forming the first layer. A subsequent binding of glucose oxidase– avidin conjugate to the target DNA and the introduction of a second layer of a redox polymer to the electrode, via layer‐by‐layer electrostatic self‐assembly, allow for electrochemical detection of the catalytic oxidation current of glucose in a PBS solution. Less than 2.0 fg of rat genomic DNA, for both regulated and house‐keeping genes, can be easily detected in 2.5 µl droplets. The proposed procedure shows very high specificity for genomic DNA in a RT–PCR mixture.</description><identifier>ISSN: 0305-1048</identifier><identifier>ISSN: 1362-4962</identifier><identifier>EISSN: 1362-4962</identifier><identifier>DOI: 10.1093/nar/gnh016</identifier><identifier>PMID: 14734816</identifier><identifier>CODEN: NARHAD</identifier><language>eng</language><publisher>England: Oxford University Press</publisher><subject>Animals ; Biotinylation ; DNA - analysis ; DNA - genetics ; DNA, Complementary - analysis ; DNA, Complementary - genetics ; Electrochemistry - methods ; Glyceraldehyde-3-Phosphate Dehydrogenases - genetics ; Liver - metabolism ; NAR Methods Online ; Nucleic Acid Hybridization ; Oligonucleotide Probes - genetics ; Oxidation-Reduction ; Polymers - chemistry ; Polymers - metabolism ; Rats ; Reverse Transcriptase Polymerase Chain Reaction ; RNA, Messenger - genetics ; RNA, Messenger - metabolism ; Sensitivity and Specificity ; Tumor Suppressor Protein p53 - genetics</subject><ispartof>Nucleic acids research, 2004-01, Vol.32 (2), p.e15-e15</ispartof><rights>Copyright Oxford University Press(England) Jan 15, 2004</rights><rights>Copyright © 2004 Oxford University Press 2004</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c500t-24603f42c17098c382585c9d45447d1ec6e35e4435526e20a4706c87dd6353b33</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC373372/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC373372/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,727,780,784,885,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/14734816$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Xie, Hong</creatorcontrib><creatorcontrib>Yu, Yuan Hong</creatorcontrib><creatorcontrib>Mao, Pei‐Lin</creatorcontrib><creatorcontrib>Gao, Zhiqiang</creatorcontrib><title>Highly sensitive amperometric detection of genomic DNA in animal tissues</title><title>Nucleic acids research</title><addtitle>Nucl. Acids Res</addtitle><description>A simple and highly sensitive method for the detection of genomic DNA in tissue samples is described. It is based on amperometric detection of target DNA by forming an analyte/polymeric activator bilayer on a gold electrode. The biotinylated target DNA is hybridized to oligonucleotide capture probes immobilized on the gold electrode, forming the first layer. A subsequent binding of glucose oxidase– avidin conjugate to the target DNA and the introduction of a second layer of a redox polymer to the electrode, via layer‐by‐layer electrostatic self‐assembly, allow for electrochemical detection of the catalytic oxidation current of glucose in a PBS solution. Less than 2.0 fg of rat genomic DNA, for both regulated and house‐keeping genes, can be easily detected in 2.5 µl droplets. The proposed procedure shows very high specificity for genomic DNA in a RT–PCR mixture.</description><subject>Animals</subject><subject>Biotinylation</subject><subject>DNA - analysis</subject><subject>DNA - genetics</subject><subject>DNA, Complementary - analysis</subject><subject>DNA, Complementary - genetics</subject><subject>Electrochemistry - methods</subject><subject>Glyceraldehyde-3-Phosphate Dehydrogenases - genetics</subject><subject>Liver - metabolism</subject><subject>NAR Methods Online</subject><subject>Nucleic Acid Hybridization</subject><subject>Oligonucleotide Probes - genetics</subject><subject>Oxidation-Reduction</subject><subject>Polymers - chemistry</subject><subject>Polymers - metabolism</subject><subject>Rats</subject><subject>Reverse Transcriptase Polymerase Chain Reaction</subject><subject>RNA, Messenger - genetics</subject><subject>RNA, Messenger - metabolism</subject><subject>Sensitivity and Specificity</subject><subject>Tumor Suppressor Protein p53 - genetics</subject><issn>0305-1048</issn><issn>1362-4962</issn><issn>1362-4962</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2004</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkU1PFEEQhjtGIwt48QeYiQcPJiPV3zMHD4jimAAe0IR46TQ9tbuNM91Ldy-Bf--Q3eDHRU6VVD1Veet9CXlJ4R2Flh8Emw4WYQlUPSEzyhWrRavYUzIDDrKmIJodspvzFQAVVIrnZIcKzUVD1Yx0nV8sh7sqY8i--Bus7LjCFEcsybuqx4Ku-BiqOK8WGOI4NT-eHVY-VDb40Q5V8TmvMe-TZ3M7ZHyxrXvk-_Gnb0ddffL185ejw5PaSYBSM6GAzwVzVEPbON4w2UjX9kIKoXuKTiGXKASXkilkYIUG5Rrd94pLfsn5Hnm_ubtaX47YOwwl2cGs0iQm3Zlovfl7EvzSLOKN4Zpzzab9N9v9FK8n3cWMPjscBhswrrNpgILSWv8XpK1SqpXNI0AmQdF76a__Aa_iOoXJLcMAZNsKLibo7QZyKeaccP7wGgVzH7eZ4jabuCf41Z9m_Ea3-U5AvQF8Lnj7MLfpp1Gaa2m6ix_m9PxYfbg470zDfwF_BrSw</recordid><startdate>20040101</startdate><enddate>20040101</enddate><creator>Xie, Hong</creator><creator>Yu, Yuan Hong</creator><creator>Mao, Pei‐Lin</creator><creator>Gao, Zhiqiang</creator><general>Oxford University Press</general><general>Oxford Publishing Limited (England)</general><scope>BSCLL</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>7QL</scope><scope>7QO</scope><scope>7QP</scope><scope>7QR</scope><scope>7SS</scope><scope>7TK</scope><scope>7TM</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>K9.</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20040101</creationdate><title>Highly sensitive amperometric detection of genomic DNA in animal tissues</title><author>Xie, Hong ; Yu, Yuan Hong ; Mao, Pei‐Lin ; Gao, Zhiqiang</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c500t-24603f42c17098c382585c9d45447d1ec6e35e4435526e20a4706c87dd6353b33</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2004</creationdate><topic>Animals</topic><topic>Biotinylation</topic><topic>DNA - analysis</topic><topic>DNA - genetics</topic><topic>DNA, Complementary - analysis</topic><topic>DNA, Complementary - genetics</topic><topic>Electrochemistry - methods</topic><topic>Glyceraldehyde-3-Phosphate Dehydrogenases - genetics</topic><topic>Liver - metabolism</topic><topic>NAR Methods Online</topic><topic>Nucleic Acid Hybridization</topic><topic>Oligonucleotide Probes - genetics</topic><topic>Oxidation-Reduction</topic><topic>Polymers - chemistry</topic><topic>Polymers - metabolism</topic><topic>Rats</topic><topic>Reverse Transcriptase Polymerase Chain Reaction</topic><topic>RNA, Messenger - genetics</topic><topic>RNA, Messenger - metabolism</topic><topic>Sensitivity and Specificity</topic><topic>Tumor Suppressor Protein p53 - genetics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Xie, Hong</creatorcontrib><creatorcontrib>Yu, Yuan Hong</creatorcontrib><creatorcontrib>Mao, Pei‐Lin</creatorcontrib><creatorcontrib>Gao, Zhiqiang</creatorcontrib><collection>Istex</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids 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>ProQuest Health & Medical Complete (Alumni)</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>PubMed Central (Full Participant titles)</collection><jtitle>Nucleic acids research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Xie, Hong</au><au>Yu, Yuan Hong</au><au>Mao, Pei‐Lin</au><au>Gao, Zhiqiang</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Highly sensitive amperometric detection of genomic DNA in animal tissues</atitle><jtitle>Nucleic acids research</jtitle><addtitle>Nucl. Acids Res</addtitle><date>2004-01-01</date><risdate>2004</risdate><volume>32</volume><issue>2</issue><spage>e15</spage><epage>e15</epage><pages>e15-e15</pages><issn>0305-1048</issn><issn>1362-4962</issn><eissn>1362-4962</eissn><coden>NARHAD</coden><abstract>A simple and highly sensitive method for the detection of genomic DNA in tissue samples is described. It is based on amperometric detection of target DNA by forming an analyte/polymeric activator bilayer on a gold electrode. The biotinylated target DNA is hybridized to oligonucleotide capture probes immobilized on the gold electrode, forming the first layer. A subsequent binding of glucose oxidase– avidin conjugate to the target DNA and the introduction of a second layer of a redox polymer to the electrode, via layer‐by‐layer electrostatic self‐assembly, allow for electrochemical detection of the catalytic oxidation current of glucose in a PBS solution. 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| subjects | Animals Biotinylation DNA - analysis DNA - genetics DNA, Complementary - analysis DNA, Complementary - genetics Electrochemistry - methods Glyceraldehyde-3-Phosphate Dehydrogenases - genetics Liver - metabolism NAR Methods Online Nucleic Acid Hybridization Oligonucleotide Probes - genetics Oxidation-Reduction Polymers - chemistry Polymers - metabolism Rats Reverse Transcriptase Polymerase Chain Reaction RNA, Messenger - genetics RNA, Messenger - metabolism Sensitivity and Specificity Tumor Suppressor Protein p53 - genetics |
| title | Highly sensitive amperometric detection of genomic DNA in animal tissues |
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