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

Karen E. Pilcher; Petra Fey; Pascale Gaudet; Anthony S. Kowal; Rex L. Chisholm

doi:10.1038/nprot.2007.180

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

Karen E. Pilcher, Petra Fey, Pascale Gaudet, Anthony S. Kowal, Rex L. Chisholm^*

^*Corresponding author for this work

FSM Office for Research

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17 Scopus citations

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.

Original language	English (US)
Pages (from-to)	1325-1328
Number of pages	4
Journal	Nature Protocols
Volume	2
Issue number	6
DOIs	https://doi.org/10.1038/nprot.2007.180
State	Published - Jun 2007

Funding

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

ASJC Scopus subject areas

General Biochemistry, Genetics and Molecular Biology

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10.1038/nprot.2007.180

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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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AU - Fey, Petra

AU - Gaudet, Pascale

AU - Kowal, Anthony S.

AU - Chisholm, Rex L.

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

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