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

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Abstract
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 1 of 2 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 were used to prepare long single strands of Drosophila ribosomal DNA (rDNA) in high yield and 42-80% pure.

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