Sodja, Ann and Davidson, Norman (1978) Gene mapping and gene enrichment by the avidin-biotin interaction: use of cytochrome-c as a polyamine bridge. Nucleic Acids Research, 5 (2). pp. 385-401. ISSN 0305-1048. http://resolver.caltech.edu/CaltechAUTHORS:SODnar78
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A modification of previously described methods of electron microscopic gene mapping and of gene enrichment based on the avidin-biotin interaction is presented. The modification consists of coupling cytochrome-c instead of pentane tane diamine to the oxidized 2', 3' terminus of an RNA by Schiff base formation and reduction. The RNA-cytochrome-c conjugate is purified by a simple chromatographic procedure; several biotins are attached to the cytochrome moiety by acylation. The extended arm between biotin and RNA gives efficient electron microscopic gene mapping of DNA:RNA-biotin hybrids with avidin-ferritin and avidin-polymethacylate sphere labels and efficient gene enrichment by buoyant banding of DNA:RNA-biotin:avidin-spheres in CsCl. A 1400 fold enrichment (thus, 25% pure) and a 90% yield of long Drosophila DNA strands with 5S RNA genes is achieved.
|Additional Information:||Copyright © 1978 Oxford University Press. Received October 31, 1977. We would like to thank Dr. Maria Pellegrini for helpful discussions on the enrichment experiments. A.S. was the recipient of a fellowship from the California Section of the American Cancer Society. This research has been supported by grant GM 10991 from the United States Public Health Service.|
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|Deposited By:||Tony Diaz|
|Deposited On:||25 Jul 2006|
|Last Modified:||26 Dec 2012 08:57|
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