{"title":"大肠杆菌DNA聚合酶I大片段超螺旋DNA测序的最佳条件","authors":"Heon M. Lim, Jacques J. Pène","doi":"10.1016/0735-0651(88)90024-6","DOIUrl":null,"url":null,"abstract":"<div><p>Sequencing ladders produced from supercoiled DNA templates with the <em>Escherichia coli</em> DNA polymerase Klenow fragment are often unreadable because of a high background and misincorporated nucleotides. This study showed that contaminating RNA molecules can interfere with template: primer hybridization. Procedures are provided for the purification of template DNA and stringent conditions for primer-template hybridization that overcome these problems.</p></div>","PeriodicalId":77714,"journal":{"name":"Gene analysis techniques","volume":"5 2","pages":"Pages 32-39"},"PeriodicalIF":0.0000,"publicationDate":"1988-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0735-0651(88)90024-6","citationCount":"53","resultStr":"{\"title\":\"Optimal conditions for supercoil DNA sequencing with the Escherichia coli DNA polymerase I large fragment\",\"authors\":\"Heon M. Lim, Jacques J. Pène\",\"doi\":\"10.1016/0735-0651(88)90024-6\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Sequencing ladders produced from supercoiled DNA templates with the <em>Escherichia coli</em> DNA polymerase Klenow fragment are often unreadable because of a high background and misincorporated nucleotides. This study showed that contaminating RNA molecules can interfere with template: primer hybridization. Procedures are provided for the purification of template DNA and stringent conditions for primer-template hybridization that overcome these problems.</p></div>\",\"PeriodicalId\":77714,\"journal\":{\"name\":\"Gene analysis techniques\",\"volume\":\"5 2\",\"pages\":\"Pages 32-39\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"1988-03-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1016/0735-0651(88)90024-6\",\"citationCount\":\"53\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Gene analysis techniques\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/0735065188900246\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Gene analysis techniques","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/0735065188900246","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
Optimal conditions for supercoil DNA sequencing with the Escherichia coli DNA polymerase I large fragment
Sequencing ladders produced from supercoiled DNA templates with the Escherichia coli DNA polymerase Klenow fragment are often unreadable because of a high background and misincorporated nucleotides. This study showed that contaminating RNA molecules can interfere with template: primer hybridization. Procedures are provided for the purification of template DNA and stringent conditions for primer-template hybridization that overcome these problems.
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