Bare Nanocapillary for DNA Separation and Genotyping Analysis in Gel-Free Solutions without Application of External Electric Field
In this work, we demonstrate DNA separation and genotyping analysis in gel-free solutions using a nanocapillary under pressure-driven conditions without application of an external electric field. The nanocapillary is a ∼50-cm-long and 500-nm-radius bare fused-silica capillary. After a DNA sample is...
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| Veröffentlicht in: | Analytical chemistry (Washington) 2008-07, Vol.80 (14), p.5583-5589 |
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| creator | Wang, Xiayan Wang, Shili Veerappan, Vijaykumar Byun, Chang Kyu Nguyen, Han Gendhar, Brina Allen, Randy D Liu, Shaorong |
| description | In this work, we demonstrate DNA separation and genotyping analysis in gel-free solutions using a nanocapillary under pressure-driven conditions without application of an external electric field. The nanocapillary is a ∼50-cm-long and 500-nm-radius bare fused-silica capillary. After a DNA sample is injected, the analytes are eluted out in a chromatographic separation format. The elution order of DNA molecules follows strictly with their sizes, with the longer DNA being eluted out faster than the shorter ones. High resolutions are obtained for both short (a few bases) and long (tens of thousands of base pairs) DNA fragments. Effects of key experimental parameters, such as eluent composition and elution pressure, on separation efficiency and resolution are investigated. We also apply this technique for DNA separations of real-world genotyping samples to demonstrate its feasibility in biological applications. PCR products (without any purification) amplified from Arabidopsis plant genomic DNA crude preparations are directly injected into the nanocapillary, and PCR-amplified DNA fragments are well resolved, allowing for unambiguous identification of samples from heterozygous and homozygous individuals. Since the capillaries used to conduct the separations are uncoated, column lifetime is virtually unlimited. The only material that is consumed in these assays is the eluent, and hence, the operation cost is low. |
| doi_str_mv | 10.1021/ac800549k |
| format | Article |
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The nanocapillary is a ∼50-cm-long and 500-nm-radius bare fused-silica capillary. After a DNA sample is injected, the analytes are eluted out in a chromatographic separation format. The elution order of DNA molecules follows strictly with their sizes, with the longer DNA being eluted out faster than the shorter ones. High resolutions are obtained for both short (a few bases) and long (tens of thousands of base pairs) DNA fragments. Effects of key experimental parameters, such as eluent composition and elution pressure, on separation efficiency and resolution are investigated. We also apply this technique for DNA separations of real-world genotyping samples to demonstrate its feasibility in biological applications. PCR products (without any purification) amplified from Arabidopsis plant genomic DNA crude preparations are directly injected into the nanocapillary, and PCR-amplified DNA fragments are well resolved, allowing for unambiguous identification of samples from heterozygous and homozygous individuals. Since the capillaries used to conduct the separations are uncoated, column lifetime is virtually unlimited. The only material that is consumed in these assays is the eluent, and hence, the operation cost is low.</description><identifier>ISSN: 0003-2700</identifier><identifier>EISSN: 1520-6882</identifier><identifier>DOI: 10.1021/ac800549k</identifier><identifier>PMID: 18500828</identifier><identifier>CODEN: ANCHAM</identifier><language>eng</language><publisher>Washington, DC: American Chemical Society</publisher><subject>Analytical biochemistry: general aspects, technics, instrumentation ; Analytical chemistry ; Analytical, structural and metabolic biochemistry ; Arabidopsis ; Arabidopsis - genetics ; Biological and medical sciences ; Biotechnology ; Buffers ; Chemistry ; Chromatographic methods and physical methods associated with chromatography ; Chromatography ; Deoxyribonucleic acid ; DNA ; DNA, Plant - genetics ; DNA, Plant - isolation & purification ; DNA, Single-Stranded - genetics ; DNA, Single-Stranded - isolation & purification ; Electric fields ; Electrons ; Electrophoresis ; Exact sciences and technology ; Fundamental and applied biological sciences. Psychology ; Gels ; Genetic engineering ; Genetic technics ; Genotype ; Genotype & phenotype ; In vitro gene amplification. Pcr technique ; Methods. Procedures. Technologies ; Nanostructures - chemistry ; Other chromatographic methods ; Pressure ; Solutions</subject><ispartof>Analytical chemistry (Washington), 2008-07, Vol.80 (14), p.5583-5589</ispartof><rights>Copyright © 2008 American Chemical Society</rights><rights>2008 INIST-CNRS</rights><rights>Copyright American Chemical Society Jul 15, 2008</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a595t-798d5dbf834e58be724e217a81d0e8d2869045b9dbe0b6904f0660183d3262633</citedby><cites>FETCH-LOGICAL-a595t-798d5dbf834e58be724e217a81d0e8d2869045b9dbe0b6904f0660183d3262633</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://pubs.acs.org/doi/pdf/10.1021/ac800549k$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/ac800549k$$EHTML$$P50$$Gacs$$H</linktohtml><link.rule.ids>230,314,780,784,885,2765,27076,27924,27925,56738,56788</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=20498237$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/18500828$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Wang, Xiayan</creatorcontrib><creatorcontrib>Wang, Shili</creatorcontrib><creatorcontrib>Veerappan, Vijaykumar</creatorcontrib><creatorcontrib>Byun, Chang Kyu</creatorcontrib><creatorcontrib>Nguyen, Han</creatorcontrib><creatorcontrib>Gendhar, Brina</creatorcontrib><creatorcontrib>Allen, Randy D</creatorcontrib><creatorcontrib>Liu, Shaorong</creatorcontrib><title>Bare Nanocapillary for DNA Separation and Genotyping Analysis in Gel-Free Solutions without Application of External Electric Field</title><title>Analytical chemistry (Washington)</title><addtitle>Anal. Chem</addtitle><description>In this work, we demonstrate DNA separation and genotyping analysis in gel-free solutions using a nanocapillary under pressure-driven conditions without application of an external electric field. The nanocapillary is a ∼50-cm-long and 500-nm-radius bare fused-silica capillary. After a DNA sample is injected, the analytes are eluted out in a chromatographic separation format. The elution order of DNA molecules follows strictly with their sizes, with the longer DNA being eluted out faster than the shorter ones. High resolutions are obtained for both short (a few bases) and long (tens of thousands of base pairs) DNA fragments. Effects of key experimental parameters, such as eluent composition and elution pressure, on separation efficiency and resolution are investigated. We also apply this technique for DNA separations of real-world genotyping samples to demonstrate its feasibility in biological applications. PCR products (without any purification) amplified from Arabidopsis plant genomic DNA crude preparations are directly injected into the nanocapillary, and PCR-amplified DNA fragments are well resolved, allowing for unambiguous identification of samples from heterozygous and homozygous individuals. Since the capillaries used to conduct the separations are uncoated, column lifetime is virtually unlimited. The only material that is consumed in these assays is the eluent, and hence, the operation cost is low.</description><subject>Analytical biochemistry: general aspects, technics, instrumentation</subject><subject>Analytical chemistry</subject><subject>Analytical, structural and metabolic biochemistry</subject><subject>Arabidopsis</subject><subject>Arabidopsis - genetics</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Buffers</subject><subject>Chemistry</subject><subject>Chromatographic methods and physical methods associated with chromatography</subject><subject>Chromatography</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>DNA, Plant - genetics</subject><subject>DNA, Plant - isolation & purification</subject><subject>DNA, Single-Stranded - genetics</subject><subject>DNA, Single-Stranded - isolation & purification</subject><subject>Electric fields</subject><subject>Electrons</subject><subject>Electrophoresis</subject><subject>Exact sciences and technology</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gels</subject><subject>Genetic engineering</subject><subject>Genetic technics</subject><subject>Genotype</subject><subject>Genotype & phenotype</subject><subject>In vitro gene amplification. Pcr technique</subject><subject>Methods. Procedures. Technologies</subject><subject>Nanostructures - chemistry</subject><subject>Other chromatographic methods</subject><subject>Pressure</subject><subject>Solutions</subject><issn>0003-2700</issn><issn>1520-6882</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2008</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNplkV1v0zAYhSMEYmVwwR9AFhJIXAReO3Hi3Exqu3YgVeOj49pyHGfz5trBTsZ6yy_HUauWjytbfh8fnfOeJHmJ4T0Ggj8IyQBoXt09SiaYEkgLxsjjZAIAWUpKgJPkWQi3ABgDLp4mJ5hRAEbYJPk1E16hS2GdFJ02Rvgtap1H55dTtFad8KLXziJhG3ShrOu3nbbXaGqF2QYdkLbx2aRLrxRaOzOMcEA_dX_jhh5Nu85ouVNwLVo89MrHn2hhlOy9lmiplWmeJ09aYYJ6sT9Pk-_LxdX8Y7r6fPFpPl2lgla0T8uKNbSpW5blirJalSRXBJeC4QYUawgrKshpXTW1gnq8t1AUgFnWZKQgRZadJmc73W6oN6qRyvZeGN55vYmpuROa_z2x-oZfu3seN1jkjEaBt3sB734MKvR8o4NUcWlWuSFwAoyyPBvB1_-At24Yk0cGl4wVjECE3u0g6V0IXrUHJxj4WCs_1BrZV39aP5L7HiPwZg-IIIVpvbBShwNHIK8YycrIpTtOh149HObC3_GizErKr76sOZvNv85W6298ddQVMhxD_G_wN4rXxho</recordid><startdate>20080715</startdate><enddate>20080715</enddate><creator>Wang, Xiayan</creator><creator>Wang, Shili</creator><creator>Veerappan, Vijaykumar</creator><creator>Byun, Chang Kyu</creator><creator>Nguyen, Han</creator><creator>Gendhar, Brina</creator><creator>Allen, Randy D</creator><creator>Liu, Shaorong</creator><general>American Chemical Society</general><scope>BSCLL</scope><scope>IQODW</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>7QF</scope><scope>7QO</scope><scope>7QQ</scope><scope>7SC</scope><scope>7SE</scope><scope>7SP</scope><scope>7SR</scope><scope>7TA</scope><scope>7TB</scope><scope>7TM</scope><scope>7U5</scope><scope>7U7</scope><scope>7U9</scope><scope>8BQ</scope><scope>8FD</scope><scope>C1K</scope><scope>F28</scope><scope>FR3</scope><scope>H8D</scope><scope>H8G</scope><scope>H94</scope><scope>JG9</scope><scope>JQ2</scope><scope>KR7</scope><scope>L7M</scope><scope>L~C</scope><scope>L~D</scope><scope>P64</scope><scope>RC3</scope><scope>5PM</scope></search><sort><creationdate>20080715</creationdate><title>Bare Nanocapillary for DNA Separation and Genotyping Analysis in Gel-Free Solutions without Application of External Electric Field</title><author>Wang, Xiayan ; Wang, Shili ; Veerappan, Vijaykumar ; Byun, Chang Kyu ; Nguyen, Han ; Gendhar, Brina ; Allen, Randy D ; Liu, Shaorong</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a595t-798d5dbf834e58be724e217a81d0e8d2869045b9dbe0b6904f0660183d3262633</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2008</creationdate><topic>Analytical biochemistry: general aspects, technics, instrumentation</topic><topic>Analytical chemistry</topic><topic>Analytical, structural and metabolic biochemistry</topic><topic>Arabidopsis</topic><topic>Arabidopsis - genetics</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>Buffers</topic><topic>Chemistry</topic><topic>Chromatographic methods and physical methods associated with chromatography</topic><topic>Chromatography</topic><topic>Deoxyribonucleic acid</topic><topic>DNA</topic><topic>DNA, Plant - genetics</topic><topic>DNA, Plant - isolation & purification</topic><topic>DNA, Single-Stranded - genetics</topic><topic>DNA, Single-Stranded - isolation & purification</topic><topic>Electric fields</topic><topic>Electrons</topic><topic>Electrophoresis</topic><topic>Exact sciences and technology</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gels</topic><topic>Genetic engineering</topic><topic>Genetic technics</topic><topic>Genotype</topic><topic>Genotype & phenotype</topic><topic>In vitro gene amplification. Pcr technique</topic><topic>Methods. Procedures. Technologies</topic><topic>Nanostructures - chemistry</topic><topic>Other chromatographic methods</topic><topic>Pressure</topic><topic>Solutions</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wang, Xiayan</creatorcontrib><creatorcontrib>Wang, Shili</creatorcontrib><creatorcontrib>Veerappan, Vijaykumar</creatorcontrib><creatorcontrib>Byun, Chang Kyu</creatorcontrib><creatorcontrib>Nguyen, Han</creatorcontrib><creatorcontrib>Gendhar, Brina</creatorcontrib><creatorcontrib>Allen, Randy D</creatorcontrib><creatorcontrib>Liu, Shaorong</creatorcontrib><collection>Istex</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Aluminium Industry Abstracts</collection><collection>Biotechnology Research Abstracts</collection><collection>Ceramic Abstracts</collection><collection>Computer and Information Systems Abstracts</collection><collection>Corrosion Abstracts</collection><collection>Electronics & Communications Abstracts</collection><collection>Engineered Materials Abstracts</collection><collection>Materials Business File</collection><collection>Mechanical & Transportation Engineering Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ANTE: Abstracts in New Technology & Engineering</collection><collection>Engineering Research Database</collection><collection>Aerospace Database</collection><collection>Copper Technical Reference Library</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Materials Research Database</collection><collection>ProQuest Computer Science Collection</collection><collection>Civil Engineering Abstracts</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>Computer and Information Systems Abstracts Academic</collection><collection>Computer and Information Systems Abstracts Professional</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Analytical chemistry (Washington)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wang, Xiayan</au><au>Wang, Shili</au><au>Veerappan, Vijaykumar</au><au>Byun, Chang Kyu</au><au>Nguyen, Han</au><au>Gendhar, Brina</au><au>Allen, Randy D</au><au>Liu, Shaorong</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Bare Nanocapillary for DNA Separation and Genotyping Analysis in Gel-Free Solutions without Application of External Electric Field</atitle><jtitle>Analytical chemistry (Washington)</jtitle><addtitle>Anal. Chem</addtitle><date>2008-07-15</date><risdate>2008</risdate><volume>80</volume><issue>14</issue><spage>5583</spage><epage>5589</epage><pages>5583-5589</pages><issn>0003-2700</issn><eissn>1520-6882</eissn><coden>ANCHAM</coden><abstract>In this work, we demonstrate DNA separation and genotyping analysis in gel-free solutions using a nanocapillary under pressure-driven conditions without application of an external electric field. The nanocapillary is a ∼50-cm-long and 500-nm-radius bare fused-silica capillary. After a DNA sample is injected, the analytes are eluted out in a chromatographic separation format. The elution order of DNA molecules follows strictly with their sizes, with the longer DNA being eluted out faster than the shorter ones. High resolutions are obtained for both short (a few bases) and long (tens of thousands of base pairs) DNA fragments. Effects of key experimental parameters, such as eluent composition and elution pressure, on separation efficiency and resolution are investigated. We also apply this technique for DNA separations of real-world genotyping samples to demonstrate its feasibility in biological applications. PCR products (without any purification) amplified from Arabidopsis plant genomic DNA crude preparations are directly injected into the nanocapillary, and PCR-amplified DNA fragments are well resolved, allowing for unambiguous identification of samples from heterozygous and homozygous individuals. Since the capillaries used to conduct the separations are uncoated, column lifetime is virtually unlimited. The only material that is consumed in these assays is the eluent, and hence, the operation cost is low.</abstract><cop>Washington, DC</cop><pub>American Chemical Society</pub><pmid>18500828</pmid><doi>10.1021/ac800549k</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Analytical biochemistry: general aspects, technics, instrumentation Analytical chemistry Analytical, structural and metabolic biochemistry Arabidopsis Arabidopsis - genetics Biological and medical sciences Biotechnology Buffers Chemistry Chromatographic methods and physical methods associated with chromatography Chromatography Deoxyribonucleic acid DNA DNA, Plant - genetics DNA, Plant - isolation & purification DNA, Single-Stranded - genetics DNA, Single-Stranded - isolation & purification Electric fields Electrons Electrophoresis Exact sciences and technology Fundamental and applied biological sciences. Psychology Gels Genetic engineering Genetic technics Genotype Genotype & phenotype In vitro gene amplification. Pcr technique Methods. Procedures. Technologies Nanostructures - chemistry Other chromatographic methods Pressure Solutions |
| title | Bare Nanocapillary for DNA Separation and Genotyping Analysis in Gel-Free Solutions without Application of External Electric Field |
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