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A reliable general purpose method for extracting genomic DNA from Dictyostelium cells

Nature Protocols volume 2, pages 1325–1328 (2007)Cite this article

Abstract

In this protocol, we present a standard method for extracting DNA from cells of the social amoeba Dictyostelium discoideum. While this procedure is similar to other phenol:chloroform-based purification methods, it is modified to account for the high level of carbohydrate and nucleases found in Dictyostelium cells. Genomic DNA can be isolated from wild-type and genetically modified cells using the described protocol, allowing molecular genetic analyses to be performed. Following cell lysis, nucleic acid extraction, and precipitation, the isolated DNA is suitable for digestion by restriction enzymes, amplification by PCR and Southern blotting. This procedure takes approximately 3 h to complete.

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Figure 1: Results from the DNA extraction protocol.

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Acknowledgements

dictyBase (http://www.dictybase.org) is supported by grants from the NIH (GM64426 and HG00022).

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Authors and Affiliations

  1. dictyBase, Center for Genetic Medicine, Northwestern University, 676 North Saint Clair Street Suite 1260, Chicago, 60611, Illinois, USA

    Karen E Pilcher, Petra Fey, Pascale Gaudet & Rex L Chisholm

  2. Center for Genetic Medicine, Northwestern University, 303 East Superior Room 7-125, Chicago, 60611, Illinois, USA

    Anthony S Kowal & Rex L Chisholm

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  1. Karen E Pilcher
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  2. Petra Fey
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  3. Pascale Gaudet
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  5. Rex L Chisholm
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Correspondence to Rex L Chisholm.

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Pilcher, K., Fey, P., Gaudet, P. et al. A reliable general purpose method for extracting genomic DNA from Dictyostelium cells. Nat Protoc 2, 1325–1328 (2007). https://doi.org/10.1038/nprot.2007.180

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