Preparation of labeled nucleic acids (nick translation and iodination) for DNA homology and rRNA hybridization experiments |
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| Authors: | Ylva M. Selin Barbara Harich J. L. Johnson |
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| Affiliation: | (1) Department of Anaerobic Microbiology, Virginia Polytechnic Institute and State University, 24061 Blacksburg, Virginia, USA |
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| Abstract: | Nick-translated DNA preparations may contain very short fragments, causing inconsistent DNA homology results. This appears to be due to extensive fragmentation of the high molecular weight DNA used in the labeling procedure. DNA preparations from organisms containing DNA of low mol% G+C content appear to be the most fragmented. The problem was circumvented by labeling these DNA preparations with125I. Sample preparation protocols have been developed for the routine125I labeling of DNA and rRNA from a wide range of organisms. |
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