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

Research output: Contribution to journalArticlepeer-review

16 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 languageEnglish (US)
Pages (from-to)1325-1328
Number of pages4
JournalNature Protocols
Volume2
Issue number6
DOIs
StatePublished - Jun 2007

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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