Mary Kay McCormick , Stylianos E Antonarakis , Philip Hieter
{"title":"琼脂糖基质中嵌入DNA的YAC克隆","authors":"Mary Kay McCormick , Stylianos E Antonarakis , Philip Hieter","doi":"10.1016/0735-0651(90)90016-9","DOIUrl":null,"url":null,"abstract":"<div><p>Yeast artificial chromosome (YAC) cloning of DNA in agarose is an alternative method to cloning from aqueous solutions. It minimizes any shearing that may result from handling of high molecular weight DNA and can be done with nanogram to microgram amounts of material, which facilitates construction of YACs from sources of DNA other than genomic DNA isolated from cells. The average size of the YACs recovered (200–1000 kb) and efficiency of transformation of ligation products (200–1000 cfu/μg) are similar to those reported using aqueous protocols. This method has been used to construct chromosome specific YACs, and it should be possible to apply the technique to the construction of chromosome specific libraries using flow sorted chromosomes as source material, and the cloning of restriction fragments isolated by preparative pulsed field gel electrophoresis.</p></div>","PeriodicalId":77714,"journal":{"name":"Gene analysis techniques","volume":"7 5","pages":"Pages 114-118"},"PeriodicalIF":0.0000,"publicationDate":"1990-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0735-0651(90)90016-9","citationCount":"5","resultStr":"{\"title\":\"YAC cloning of DNA embedded in an agarose matrix\",\"authors\":\"Mary Kay McCormick , Stylianos E Antonarakis , Philip Hieter\",\"doi\":\"10.1016/0735-0651(90)90016-9\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Yeast artificial chromosome (YAC) cloning of DNA in agarose is an alternative method to cloning from aqueous solutions. It minimizes any shearing that may result from handling of high molecular weight DNA and can be done with nanogram to microgram amounts of material, which facilitates construction of YACs from sources of DNA other than genomic DNA isolated from cells. The average size of the YACs recovered (200–1000 kb) and efficiency of transformation of ligation products (200–1000 cfu/μg) are similar to those reported using aqueous protocols. This method has been used to construct chromosome specific YACs, and it should be possible to apply the technique to the construction of chromosome specific libraries using flow sorted chromosomes as source material, and the cloning of restriction fragments isolated by preparative pulsed field gel electrophoresis.</p></div>\",\"PeriodicalId\":77714,\"journal\":{\"name\":\"Gene analysis techniques\",\"volume\":\"7 5\",\"pages\":\"Pages 114-118\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"1990-09-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1016/0735-0651(90)90016-9\",\"citationCount\":\"5\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Gene analysis techniques\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/0735065190900169\",\"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/0735065190900169","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
Yeast artificial chromosome (YAC) cloning of DNA in agarose is an alternative method to cloning from aqueous solutions. It minimizes any shearing that may result from handling of high molecular weight DNA and can be done with nanogram to microgram amounts of material, which facilitates construction of YACs from sources of DNA other than genomic DNA isolated from cells. The average size of the YACs recovered (200–1000 kb) and efficiency of transformation of ligation products (200–1000 cfu/μg) are similar to those reported using aqueous protocols. This method has been used to construct chromosome specific YACs, and it should be possible to apply the technique to the construction of chromosome specific libraries using flow sorted chromosomes as source material, and the cloning of restriction fragments isolated by preparative pulsed field gel electrophoresis.
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