Comparison between Pulsed-field and Constant-field Gel Electrophoresis for Measurement of DNA Double-strand Breaks in Irradiated Chinese Hamster Ovary Cells
Summary Pulsed-field gel electrophoresis (PFGE) is one of the most sensitive methods for detecting DNA double-strand breaks in mammalian cells. However, it has been observed that constant-field gel electrophoresis (CFGE), when optimized, can detect breaks with equal efficiency. The migration of DNA...
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| Veröffentlicht in: | International journal of radiation biology 1991, Vol.60 (5), p.779-790 |
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| container_title | International journal of radiation biology |
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| creator | Wlodek, D. Banáth, J. Olive, P.L. |
| description | Summary
Pulsed-field gel electrophoresis (PFGE) is one of the most sensitive methods for detecting DNA double-strand breaks in mammalian cells. However, it has been observed that constant-field gel electrophoresis (CFGE), when optimized, can detect breaks with equal efficiency. The migration of DNA from the well and the separation of DNA molecules according to size appear to be different processes; only the latter requires the application of PFGE. CFGE is very sensitive and can detect DNA damage produced by less than 5 Gy of radiation. Low voltage (ca. 0·6 V/cm) during electrophoresis appears to be essential for the migration of the largest fraction of DNA from the agarose plug containing the cells; the electrophoresis run time, cell density in the plug, agarose concentration, nature of detergent and extent of radiolabelling are less important. It is concluded that CFGE is equally sensitive but more rapid and economical than PFGE for the measurement of DNA damage. |
| doi_str_mv | 10.1080/09553009114552591 |
| format | Article |
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Pulsed-field gel electrophoresis (PFGE) is one of the most sensitive methods for detecting DNA double-strand breaks in mammalian cells. However, it has been observed that constant-field gel electrophoresis (CFGE), when optimized, can detect breaks with equal efficiency. The migration of DNA from the well and the separation of DNA molecules according to size appear to be different processes; only the latter requires the application of PFGE. CFGE is very sensitive and can detect DNA damage produced by less than 5 Gy of radiation. Low voltage (ca. 0·6 V/cm) during electrophoresis appears to be essential for the migration of the largest fraction of DNA from the agarose plug containing the cells; the electrophoresis run time, cell density in the plug, agarose concentration, nature of detergent and extent of radiolabelling are less important. It is concluded that CFGE is equally sensitive but more rapid and economical than PFGE for the measurement of DNA damage.</description><identifier>ISSN: 0955-3002</identifier><identifier>EISSN: 1362-3095</identifier><identifier>DOI: 10.1080/09553009114552591</identifier><identifier>PMID: 1680949</identifier><language>eng</language><publisher>London: Informa UK Ltd</publisher><subject>Animals ; Biological and medical sciences ; CHO Cells - radiation effects ; Cricetinae ; DNA - radiation effects ; DNA Damage ; Dose-Response Relationship, Radiation ; Electrophoresis, Agar Gel ; Electrophoresis, Gel, Pulsed-Field ; Fundamental and applied biological sciences. Psychology ; Tissues, organs and organisms biophysics</subject><ispartof>International journal of radiation biology, 1991, Vol.60 (5), p.779-790</ispartof><rights>1991 Informa UK Ltd All rights reserved: reproduction in whole or part not permitted 1991</rights><rights>1992 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c496t-3d6ccec25e16c55ccdce313855d70fbdaa19e05177567af6f1ab2d59bd3d2e553</citedby><cites>FETCH-LOGICAL-c496t-3d6ccec25e16c55ccdce313855d70fbdaa19e05177567af6f1ab2d59bd3d2e553</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.tandfonline.com/doi/pdf/10.1080/09553009114552591$$EPDF$$P50$$Ginformaworld$$H</linktopdf><linktohtml>$$Uhttps://www.tandfonline.com/doi/full/10.1080/09553009114552591$$EHTML$$P50$$Ginformaworld$$H</linktohtml><link.rule.ids>314,777,781,4010,27904,27905,27906,59626,59732,60415,60521,61200,61235,61381,61416</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=5129529$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1680949$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Wlodek, D.</creatorcontrib><creatorcontrib>Banáth, J.</creatorcontrib><creatorcontrib>Olive, P.L.</creatorcontrib><title>Comparison between Pulsed-field and Constant-field Gel Electrophoresis for Measurement of DNA Double-strand Breaks in Irradiated Chinese Hamster Ovary Cells</title><title>International journal of radiation biology</title><addtitle>Int J Radiat Biol</addtitle><description>Summary
Pulsed-field gel electrophoresis (PFGE) is one of the most sensitive methods for detecting DNA double-strand breaks in mammalian cells. However, it has been observed that constant-field gel electrophoresis (CFGE), when optimized, can detect breaks with equal efficiency. The migration of DNA from the well and the separation of DNA molecules according to size appear to be different processes; only the latter requires the application of PFGE. CFGE is very sensitive and can detect DNA damage produced by less than 5 Gy of radiation. Low voltage (ca. 0·6 V/cm) during electrophoresis appears to be essential for the migration of the largest fraction of DNA from the agarose plug containing the cells; the electrophoresis run time, cell density in the plug, agarose concentration, nature of detergent and extent of radiolabelling are less important. It is concluded that CFGE is equally sensitive but more rapid and economical than PFGE for the measurement of DNA damage.</description><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>CHO Cells - radiation effects</subject><subject>Cricetinae</subject><subject>DNA - radiation effects</subject><subject>DNA Damage</subject><subject>Dose-Response Relationship, Radiation</subject><subject>Electrophoresis, Agar Gel</subject><subject>Electrophoresis, Gel, Pulsed-Field</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Tissues, organs and organisms biophysics</subject><issn>0955-3002</issn><issn>1362-3095</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1991</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kM1uFDEQhC0ECkvgATgg-cB1iD1ez64FlzD5lQLhAOdRj93WOnjsle0hyrvwsHi1Cwgh5eSWq6pV_RHymrN3nK3ZCVNSCsYU50spW6n4E7LgomsbUZWnZLHT68za5-RFznesTkysj8gR79ZMLdWC_OzjtIXkcgx0xHKPGOiX2Wc0jXXoDYVgaB9DLhDK4esSPT33qEuK201MmF2mNib6CSHPCScMhUZLzz6f0rM4jx6bXNJuz8eE8D1TF-h1SmAcFKzLNy5gRnoFUy6Y6O0PSA-0R-_zS_LMQu3y6vAek28X51_7q-bm9vK6P71p9FJ1pRGm0xp1K5F3WkqtjUbBxVpKs2J2NABcIZN8tZLdCmxnOYytkWo0wrRYCR4Tvt-rU8w5oR22yU21xsDZsAM9_Ae6Zt7sM9t5nND8TezJVv3tQYeswdsKQLv8xyZ5q2S7s33Y21yoCCe4j8mbocCDj-l3RjzW4v0_8Q2CLxsNCYe7OKdQqT1ywy8NYq2d</recordid><startdate>1991</startdate><enddate>1991</enddate><creator>Wlodek, D.</creator><creator>Banáth, J.</creator><creator>Olive, P.L.</creator><general>Informa UK Ltd</general><general>Taylor & Francis</general><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></search><sort><creationdate>1991</creationdate><title>Comparison between Pulsed-field and Constant-field Gel Electrophoresis for Measurement of DNA Double-strand Breaks in Irradiated Chinese Hamster Ovary Cells</title><author>Wlodek, D. ; Banáth, J. ; Olive, P.L.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c496t-3d6ccec25e16c55ccdce313855d70fbdaa19e05177567af6f1ab2d59bd3d2e553</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1991</creationdate><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>CHO Cells - radiation effects</topic><topic>Cricetinae</topic><topic>DNA - radiation effects</topic><topic>DNA Damage</topic><topic>Dose-Response Relationship, Radiation</topic><topic>Electrophoresis, Agar Gel</topic><topic>Electrophoresis, Gel, Pulsed-Field</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Tissues, organs and organisms biophysics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wlodek, D.</creatorcontrib><creatorcontrib>Banáth, J.</creatorcontrib><creatorcontrib>Olive, P.L.</creatorcontrib><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><jtitle>International journal of radiation biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wlodek, D.</au><au>Banáth, J.</au><au>Olive, P.L.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Comparison between Pulsed-field and Constant-field Gel Electrophoresis for Measurement of DNA Double-strand Breaks in Irradiated Chinese Hamster Ovary Cells</atitle><jtitle>International journal of radiation biology</jtitle><addtitle>Int J Radiat Biol</addtitle><date>1991</date><risdate>1991</risdate><volume>60</volume><issue>5</issue><spage>779</spage><epage>790</epage><pages>779-790</pages><issn>0955-3002</issn><eissn>1362-3095</eissn><abstract>Summary
Pulsed-field gel electrophoresis (PFGE) is one of the most sensitive methods for detecting DNA double-strand breaks in mammalian cells. However, it has been observed that constant-field gel electrophoresis (CFGE), when optimized, can detect breaks with equal efficiency. The migration of DNA from the well and the separation of DNA molecules according to size appear to be different processes; only the latter requires the application of PFGE. CFGE is very sensitive and can detect DNA damage produced by less than 5 Gy of radiation. Low voltage (ca. 0·6 V/cm) during electrophoresis appears to be essential for the migration of the largest fraction of DNA from the agarose plug containing the cells; the electrophoresis run time, cell density in the plug, agarose concentration, nature of detergent and extent of radiolabelling are less important. It is concluded that CFGE is equally sensitive but more rapid and economical than PFGE for the measurement of DNA damage.</abstract><cop>London</cop><pub>Informa UK Ltd</pub><pmid>1680949</pmid><doi>10.1080/09553009114552591</doi><tpages>12</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0955-3002 |
| ispartof | International journal of radiation biology, 1991, Vol.60 (5), p.779-790 |
| issn | 0955-3002 1362-3095 |
| language | eng |
| recordid | cdi_crossref_primary_10_1080_09553009114552591 |
| source | MEDLINE; Taylor & Francis:Master (3349 titles); Taylor & Francis Medical Library - CRKN |
| subjects | Animals Biological and medical sciences CHO Cells - radiation effects Cricetinae DNA - radiation effects DNA Damage Dose-Response Relationship, Radiation Electrophoresis, Agar Gel Electrophoresis, Gel, Pulsed-Field Fundamental and applied biological sciences. Psychology Tissues, organs and organisms biophysics |
| title | Comparison between Pulsed-field and Constant-field Gel Electrophoresis for Measurement of DNA Double-strand Breaks in Irradiated Chinese Hamster Ovary Cells |
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