A method for gene enrichment based on the avidin-biotin interaction. Application to the Drosophila ribosomal RNA genes

作者：Jerry E. Manning, Maria Pellegrini, Norman Davidson · 发表于：Biochemistry · 年份：1977 · DOI：10.1021/bi00626a020 · 被引用次数：40 · 研究领域：Biotin and Related Studies、RNA and protein synthesis mechanisms、Click Chemistry and Applications

A method of enriching, from the total DNA of an organism, for long DNA strands carrying a particular gene is described. The purified RNA corresponding to the gene is covalently attached to biotin via a cytochrome c bridge. This modified RNA is hybridized to the total DNA. Those DNA strands which hybridize are separated from all the other DNA, using the avidin-biotin interaction, by one of two methods. Avidin is covalently attached to submicroscopic polymer spheres; the complexes of avidin spheres with the DNA: RNA-biotin hybrids band in CsCl at a much lower buoyant density than does free DNA. Alternatively, the DNA:RNA-biotin hybrids are isolated by affinity chromatography on an avidin-solid support column. These methods have been used to prepare long single strands of Drosophila ribosomal DNA (rDNA) in high yield and 42 to 80% pure.