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A general purpose method for extracting RNA from Dictyostelium cells

Abstract

Here we present a protocol for the extraction of RNA from Dictyostelium discoideum. Dictyostelium is a social amoeba that undergoes a basic developmental program, and therefore analysis of RNA levels over a time course is a commonly used technique. This procedure is similar to other guanidine thiocyanate-based methods; however, it has been adjusted because of the large quantities of carbohydrate and nucleases found in Dictyostelium cells. After cell lysis and phenol:chloroform extraction, the resulting high-quality RNA isolated with the described protocol allows the molecular genetic analysis of wild-type and genetically modified cells. The purified RNA can be used for analyses such as northern blotting, RT-PCR and microarrays. This procedure requires approximately 2 h to complete.

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Figure 1: RNA isolated from D. discoideum.

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Acknowledgements

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

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Correspondence to Rex L Chisholm.

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Pilcher, K., Gaudet, P., Fey, P. et al. A general purpose method for extracting RNA from Dictyostelium cells. Nat Protoc 2, 1329–1332 (2007). https://doi.org/10.1038/nprot.2007.191

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