Separation of 1-23-kb complementary DNA strands by urea-agarose gel electrophoresis
Double-stranded (ds), as well as denatured, single-stranded (ss) DNA samples can be analyzed on urea-agarose gels. Here we report that after denaturation by heat in the presence of 8 M urea, the two strands of the same ds DNA fragment of ~1-20-kb size migrate differently in 1 M urea containing agaro...
Gespeichert in:
| Veröffentlicht in: | Nucleic acids research 2009-09, Vol.37 (17), p.e112-e112 |
|---|---|
| Hauptverfasser: | , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | e112 |
|---|---|
| container_issue | 17 |
| container_start_page | e112 |
| container_title | Nucleic acids research |
| container_volume | 37 |
| creator | Hegedüs, Éva Kókai, Endre Kotlyar, Alexander Dombrádi, Viktor Szabó, Gábor |
| description | Double-stranded (ds), as well as denatured, single-stranded (ss) DNA samples can be analyzed on urea-agarose gels. Here we report that after denaturation by heat in the presence of 8 M urea, the two strands of the same ds DNA fragment of ~1-20-kb size migrate differently in 1 M urea containing agarose gels. The two strands are readily distinguished on Southern blots by ss-specific probes. The different migration of the two strands could be attributed to their different, base composition-dependent conformation impinging on the electrophoretic mobility of the ss molecules. This phenomenon can be exploited for the efficient preparation of strand-specific probes and for the separation of the complementary DNA strands for subsequent analysis, offering a new tool for various cell biological research areas. |
| doi_str_mv | 10.1093/nar/gkp539 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_2761254</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><oup_id>10.1093/nar/gkp539</oup_id><sourcerecordid>734085204</sourcerecordid><originalsourceid>FETCH-LOGICAL-c500t-a313948c15890acbc5ccded116e30c07541b1b9845167ad0669a705806447d8b3</originalsourceid><addsrcrecordid>eNp90Utv1DAUBWALgegwsOEHQDaAhBR6r5_JBqmUxyCNYJFWQmwsx_FMwyRxsBNE_z2uMiqw6coLfzo69iHkKcIbhJKdDiac7g-jYOU9skImac5LSe-TFTAQOQIvTsijGH8AIEfBH5ITLIVgWJQrUlVuNMFMrR8yv8swpyw_1Jn1_di53g2TCdfZ-y9nWZyCGZqY1dfZHJzJzd4EH122d13mOmen4McrH1xs42PyYGe66J4czzW5_Pjh4nyTb79--nx-ts2tAJhyw5CVvLAoihKMra2wtnENonQMLCjBsca6LLhAqUwDUpZGgShAcq6aomZr8nbJHee6d41NbYPp9BjaPrXW3rT6_5uhvdJ7_0tTJZEKngJeHQOC_zm7OOm-jdZ1nRmcn6NWjEMhKNzIl3dKmlpTUJDg6wXa9DsxuN1tHQR9s5ZOa-llrYSf_fuAv_Q4TwIvFuDn8e6gfHFtnNzvW2nCQUvFlNCbb9_1RVW821S4Tc9ak-eL3xmvzT60UV9WFJABypIiKPYH7WC1ug</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>21162070</pqid></control><display><type>article</type><title>Separation of 1-23-kb complementary DNA strands by urea-agarose gel electrophoresis</title><source>MEDLINE</source><source>DOAJ Directory of Open Access Journals</source><source>Oxford Open</source><source>PubMed Central</source><source>Free Full-Text Journals in Chemistry</source><creator>Hegedüs, Éva ; Kókai, Endre ; Kotlyar, Alexander ; Dombrádi, Viktor ; Szabó, Gábor</creator><creatorcontrib>Hegedüs, Éva ; Kókai, Endre ; Kotlyar, Alexander ; Dombrádi, Viktor ; Szabó, Gábor</creatorcontrib><description>Double-stranded (ds), as well as denatured, single-stranded (ss) DNA samples can be analyzed on urea-agarose gels. Here we report that after denaturation by heat in the presence of 8 M urea, the two strands of the same ds DNA fragment of ~1-20-kb size migrate differently in 1 M urea containing agarose gels. The two strands are readily distinguished on Southern blots by ss-specific probes. The different migration of the two strands could be attributed to their different, base composition-dependent conformation impinging on the electrophoretic mobility of the ss molecules. This phenomenon can be exploited for the efficient preparation of strand-specific probes and for the separation of the complementary DNA strands for subsequent analysis, offering a new tool for various cell biological research areas.</description><identifier>ISSN: 0305-1048</identifier><identifier>EISSN: 1362-4962</identifier><identifier>DOI: 10.1093/nar/gkp539</identifier><identifier>PMID: 19553189</identifier><language>eng</language><publisher>England: Oxford University Press</publisher><subject>DNA, Complementary - chemistry ; DNA, Complementary - isolation & purification ; DNA, Fungal - chemistry ; DNA, Fungal - isolation & purification ; DNA, Single-Stranded - isolation & purification ; Electrophoresis, Agar Gel - methods ; Methods Online ; Nucleic Acid Denaturation ; Saccharomyces cerevisiae - genetics ; Urea - chemistry</subject><ispartof>Nucleic acids research, 2009-09, Vol.37 (17), p.e112-e112</ispartof><rights>2009 The Author(s) 2009</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c500t-a313948c15890acbc5ccded116e30c07541b1b9845167ad0669a705806447d8b3</citedby><cites>FETCH-LOGICAL-c500t-a313948c15890acbc5ccded116e30c07541b1b9845167ad0669a705806447d8b3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2761254/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2761254/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,860,881,1598,27901,27902,53766,53768</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/19553189$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Hegedüs, Éva</creatorcontrib><creatorcontrib>Kókai, Endre</creatorcontrib><creatorcontrib>Kotlyar, Alexander</creatorcontrib><creatorcontrib>Dombrádi, Viktor</creatorcontrib><creatorcontrib>Szabó, Gábor</creatorcontrib><title>Separation of 1-23-kb complementary DNA strands by urea-agarose gel electrophoresis</title><title>Nucleic acids research</title><addtitle>Nucleic Acids Res</addtitle><description>Double-stranded (ds), as well as denatured, single-stranded (ss) DNA samples can be analyzed on urea-agarose gels. Here we report that after denaturation by heat in the presence of 8 M urea, the two strands of the same ds DNA fragment of ~1-20-kb size migrate differently in 1 M urea containing agarose gels. The two strands are readily distinguished on Southern blots by ss-specific probes. The different migration of the two strands could be attributed to their different, base composition-dependent conformation impinging on the electrophoretic mobility of the ss molecules. This phenomenon can be exploited for the efficient preparation of strand-specific probes and for the separation of the complementary DNA strands for subsequent analysis, offering a new tool for various cell biological research areas.</description><subject>DNA, Complementary - chemistry</subject><subject>DNA, Complementary - isolation & purification</subject><subject>DNA, Fungal - chemistry</subject><subject>DNA, Fungal - isolation & purification</subject><subject>DNA, Single-Stranded - isolation & purification</subject><subject>Electrophoresis, Agar Gel - methods</subject><subject>Methods Online</subject><subject>Nucleic Acid Denaturation</subject><subject>Saccharomyces cerevisiae - genetics</subject><subject>Urea - chemistry</subject><issn>0305-1048</issn><issn>1362-4962</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2009</creationdate><recordtype>article</recordtype><sourceid>TOX</sourceid><sourceid>EIF</sourceid><recordid>eNp90Utv1DAUBWALgegwsOEHQDaAhBR6r5_JBqmUxyCNYJFWQmwsx_FMwyRxsBNE_z2uMiqw6coLfzo69iHkKcIbhJKdDiac7g-jYOU9skImac5LSe-TFTAQOQIvTsijGH8AIEfBH5ITLIVgWJQrUlVuNMFMrR8yv8swpyw_1Jn1_di53g2TCdfZ-y9nWZyCGZqY1dfZHJzJzd4EH122d13mOmen4McrH1xs42PyYGe66J4czzW5_Pjh4nyTb79--nx-ts2tAJhyw5CVvLAoihKMra2wtnENonQMLCjBsca6LLhAqUwDUpZGgShAcq6aomZr8nbJHee6d41NbYPp9BjaPrXW3rT6_5uhvdJ7_0tTJZEKngJeHQOC_zm7OOm-jdZ1nRmcn6NWjEMhKNzIl3dKmlpTUJDg6wXa9DsxuN1tHQR9s5ZOa-llrYSf_fuAv_Q4TwIvFuDn8e6gfHFtnNzvW2nCQUvFlNCbb9_1RVW821S4Tc9ak-eL3xmvzT60UV9WFJABypIiKPYH7WC1ug</recordid><startdate>20090901</startdate><enddate>20090901</enddate><creator>Hegedüs, Éva</creator><creator>Kókai, Endre</creator><creator>Kotlyar, Alexander</creator><creator>Dombrádi, Viktor</creator><creator>Szabó, Gábor</creator><general>Oxford University Press</general><scope>FBQ</scope><scope>BSCLL</scope><scope>TOX</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>7TM</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20090901</creationdate><title>Separation of 1-23-kb complementary DNA strands by urea-agarose gel electrophoresis</title><author>Hegedüs, Éva ; Kókai, Endre ; Kotlyar, Alexander ; Dombrádi, Viktor ; Szabó, Gábor</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c500t-a313948c15890acbc5ccded116e30c07541b1b9845167ad0669a705806447d8b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2009</creationdate><topic>DNA, Complementary - chemistry</topic><topic>DNA, Complementary - isolation & purification</topic><topic>DNA, Fungal - chemistry</topic><topic>DNA, Fungal - isolation & purification</topic><topic>DNA, Single-Stranded - isolation & purification</topic><topic>Electrophoresis, Agar Gel - methods</topic><topic>Methods Online</topic><topic>Nucleic Acid Denaturation</topic><topic>Saccharomyces cerevisiae - genetics</topic><topic>Urea - chemistry</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Hegedüs, Éva</creatorcontrib><creatorcontrib>Kókai, Endre</creatorcontrib><creatorcontrib>Kotlyar, Alexander</creatorcontrib><creatorcontrib>Dombrádi, Viktor</creatorcontrib><creatorcontrib>Szabó, Gábor</creatorcontrib><collection>AGRIS</collection><collection>Istex</collection><collection>Oxford Open</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Nucleic Acids Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</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>Hegedüs, Éva</au><au>Kókai, Endre</au><au>Kotlyar, Alexander</au><au>Dombrádi, Viktor</au><au>Szabó, Gábor</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Separation of 1-23-kb complementary DNA strands by urea-agarose gel electrophoresis</atitle><jtitle>Nucleic acids research</jtitle><addtitle>Nucleic Acids Res</addtitle><date>2009-09-01</date><risdate>2009</risdate><volume>37</volume><issue>17</issue><spage>e112</spage><epage>e112</epage><pages>e112-e112</pages><issn>0305-1048</issn><eissn>1362-4962</eissn><abstract>Double-stranded (ds), as well as denatured, single-stranded (ss) DNA samples can be analyzed on urea-agarose gels. Here we report that after denaturation by heat in the presence of 8 M urea, the two strands of the same ds DNA fragment of ~1-20-kb size migrate differently in 1 M urea containing agarose gels. The two strands are readily distinguished on Southern blots by ss-specific probes. The different migration of the two strands could be attributed to their different, base composition-dependent conformation impinging on the electrophoretic mobility of the ss molecules. This phenomenon can be exploited for the efficient preparation of strand-specific probes and for the separation of the complementary DNA strands for subsequent analysis, offering a new tool for various cell biological research areas.</abstract><cop>England</cop><pub>Oxford University Press</pub><pmid>19553189</pmid><doi>10.1093/nar/gkp539</doi><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0305-1048 |
| ispartof | Nucleic acids research, 2009-09, Vol.37 (17), p.e112-e112 |
| issn | 0305-1048 1362-4962 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_2761254 |
| source | MEDLINE; DOAJ Directory of Open Access Journals; Oxford Open; PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | DNA, Complementary - chemistry DNA, Complementary - isolation & purification DNA, Fungal - chemistry DNA, Fungal - isolation & purification DNA, Single-Stranded - isolation & purification Electrophoresis, Agar Gel - methods Methods Online Nucleic Acid Denaturation Saccharomyces cerevisiae - genetics Urea - chemistry |
| title | Separation of 1-23-kb complementary DNA strands by urea-agarose gel electrophoresis |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-07T19%3A33%3A30IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Separation%20of%201-23-kb%20complementary%20DNA%20strands%20by%20urea-agarose%20gel%20electrophoresis&rft.jtitle=Nucleic%20acids%20research&rft.au=Heged%C3%BCs,%20%C3%89va&rft.date=2009-09-01&rft.volume=37&rft.issue=17&rft.spage=e112&rft.epage=e112&rft.pages=e112-e112&rft.issn=0305-1048&rft.eissn=1362-4962&rft_id=info:doi/10.1093/nar/gkp539&rft_dat=%3Cproquest_pubme%3E734085204%3C/proquest_pubme%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=21162070&rft_id=info:pmid/19553189&rft_oup_id=10.1093/nar/gkp539&rfr_iscdi=true |