A buoyant method for the determination of the superhelix density of closed circular DNA
A convenient method for the determination of the superhelix density of a closed circular DNA has been developed and tested with a series of closed DNA's with different superhelix densities and molecular weights. The method is based on the superhelix density-dependent binding affinity of closed...
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| Veröffentlicht in: | Journal of molecular biology 1971-11, Vol.62 (1), p.1-19 |
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| container_title | Journal of molecular biology |
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| creator | Gray, Horace B. Upholt, William B. Vinograd, Jerome |
| description | A convenient method for the determination of the superhelix density of a closed circular DNA has been developed and tested with a series of closed DNA's with different superhelix densities and molecular weights. The method is based on the superhelix density-dependent binding affinity of closed DNA for the intercalating dye, ethidium bromide. The superhelix density is calculated from the observed separation between nicked and closed DNA in a cesium chlorideethidium bromide density gradient centrifuged under specified conditions in the preparative ultracentrifuge. The linear relation between the separations and the superhelix density obtained in this study is in good agreement with that predicted by Bauer & Vinograd (1970b). |
| doi_str_mv | 10.1016/0022-2836(71)90127-6 |
| format | Article |
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The method is based on the superhelix density-dependent binding affinity of closed DNA for the intercalating dye, ethidium bromide. The superhelix density is calculated from the observed separation between nicked and closed DNA in a cesium chlorideethidium bromide density gradient centrifuged under specified conditions in the preparative ultracentrifuge. 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The method is based on the superhelix density-dependent binding affinity of closed DNA for the intercalating dye, ethidium bromide. The superhelix density is calculated from the observed separation between nicked and closed DNA in a cesium chlorideethidium bromide density gradient centrifuged under specified conditions in the preparative ultracentrifuge. The linear relation between the separations and the superhelix density obtained in this study is in good agreement with that predicted by Bauer & Vinograd (1970b).</description><subject>Binding Sites</subject><subject>Centrifugation, Density Gradient</subject><subject>DNA, Viral</subject><subject>Molecular Biology</subject><subject>Molecular Weight</subject><subject>Nucleic Acid Conformation</subject><subject>Quaternary Ammonium Compounds</subject><subject>Quinolines</subject><subject>Simian virus 40</subject><subject>Spectrophotometry</subject><subject>Ultracentrifugation</subject><issn>0022-2836</issn><issn>1089-8638</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1971</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kE9LxDAQxYMouq5-A4WeRA_VTJKm7UVY_A-iF8VjaNMJG2mbNUnF_fZ23WWPnmaY92aG9yPkBOglUJBXlDKWsoLL8xwuSgosT-UOmQAtyrSQvNglk63lgByG8Ekpzbgo9sm-4JxL4BPyMUvqwS2rPiYdxrlrEuN8EueYNBjRd7avonV94szfMAwL9HNs7c-o98HG5UrRrQvYJNp6PbSVT25fZkdkz1RtwONNnZL3-7u3m8f0-fXh6Wb2nGrBeEzrjAIakbMCDEKmJUhaSo6S55zVHIUsWSHKcmyrjMpajLlExdCgBmDG8Ck5W99dePc1YIiqs0Fj21Y9uiGoApjIZQ6jUayN2rsQPBq18Lar_FIBVSueagVLrWCpHNQfTyXHtdPN_aHusNkubQCO-vVaxzHkt0WvgrbYa2ysRx1V4-z_D34BK3iDcA</recordid><startdate>19711128</startdate><enddate>19711128</enddate><creator>Gray, Horace B.</creator><creator>Upholt, William B.</creator><creator>Vinograd, Jerome</creator><general>Elsevier Ltd</general><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></search><sort><creationdate>19711128</creationdate><title>A buoyant method for the determination of the superhelix density of closed circular DNA</title><author>Gray, Horace B. ; Upholt, William B. ; Vinograd, Jerome</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c423t-b501ef47281fe15c6160963e63732b3e46928499b3ea506b46384a2efec112ff3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1971</creationdate><topic>Binding Sites</topic><topic>Centrifugation, Density Gradient</topic><topic>DNA, Viral</topic><topic>Molecular Biology</topic><topic>Molecular Weight</topic><topic>Nucleic Acid Conformation</topic><topic>Quaternary Ammonium Compounds</topic><topic>Quinolines</topic><topic>Simian virus 40</topic><topic>Spectrophotometry</topic><topic>Ultracentrifugation</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Gray, Horace B.</creatorcontrib><creatorcontrib>Upholt, William B.</creatorcontrib><creatorcontrib>Vinograd, Jerome</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><jtitle>Journal of molecular biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Gray, Horace B.</au><au>Upholt, William B.</au><au>Vinograd, Jerome</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A buoyant method for the determination of the superhelix density of closed circular DNA</atitle><jtitle>Journal of molecular biology</jtitle><addtitle>J Mol Biol</addtitle><date>1971-11-28</date><risdate>1971</risdate><volume>62</volume><issue>1</issue><spage>1</spage><epage>19</epage><pages>1-19</pages><issn>0022-2836</issn><eissn>1089-8638</eissn><abstract>A convenient method for the determination of the superhelix density of a closed circular DNA has been developed and tested with a series of closed DNA's with different superhelix densities and molecular weights. The method is based on the superhelix density-dependent binding affinity of closed DNA for the intercalating dye, ethidium bromide. The superhelix density is calculated from the observed separation between nicked and closed DNA in a cesium chlorideethidium bromide density gradient centrifuged under specified conditions in the preparative ultracentrifuge. The linear relation between the separations and the superhelix density obtained in this study is in good agreement with that predicted by Bauer & Vinograd (1970b).</abstract><cop>England</cop><pub>Elsevier Ltd</pub><pmid>4333613</pmid><doi>10.1016/0022-2836(71)90127-6</doi><tpages>19</tpages></addata></record> |
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| ispartof | Journal of molecular biology, 1971-11, Vol.62 (1), p.1-19 |
| issn | 0022-2836 1089-8638 |
| language | eng |
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| source | MEDLINE; Elsevier ScienceDirect Journals Complete |
| subjects | Binding Sites Centrifugation, Density Gradient DNA, Viral Molecular Biology Molecular Weight Nucleic Acid Conformation Quaternary Ammonium Compounds Quinolines Simian virus 40 Spectrophotometry Ultracentrifugation |
| title | A buoyant method for the determination of the superhelix density of closed circular DNA |
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