Abstract
The isolation of chloroplast DNA fromChlamydomonas reinhardtii requires the efficient separation of this AT-rich genome from the GC-rich nuclear genome by density-gradient centrifugation. We describe a simple and efficient method for separating these DNA fractions by using a sodium iodide gradient in combination with the DNA-binding dye, bisbenzimide. The yield of chloroplast DNA is close to the theoretical maximum and the DNA is suitable for restriction enzyme analysis and cloning. This method is applicable to the isolation of AT-rich plastid genomes from other organisms and may be appropriate as a general method for separating species of DNA that differ in their AT/GC ratios.
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Abbreviations
- EtBr:
-
ethidium bromide
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An erratum to this article is available at http://dx.doi.org/10.1007/BF02905342.
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O'connor, H.E., Stevens, D.R., Ruffle, S.V. et al. Am improved procedure for the isolation of chloroplast DNA fromChlamydomonas reinhardtii . Plant Mol Biol Rep 11, 207–211 (1993). https://doi.org/10.1007/BF02669846
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DOI: https://doi.org/10.1007/BF02669846