A general purpose method for extracting RNA from Dictyostelium cells

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

doi:10.1038/nprot.2007.191

A general purpose method for extracting RNA from Dictyostelium cells

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

^*Corresponding author for this work

FSM Office for Research

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10 Scopus citations

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.

Original language	English (US)
Pages (from-to)	1329-1332
Number of pages	4
Journal	Nature Protocols
Volume	2
Issue number	6
DOIs	https://doi.org/10.1038/nprot.2007.191
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.2007.191

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title = "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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AU - Gaudet, Pascale

AU - Fey, Petra

AU - Kowal, Anthony S.

AU - Chisholm, Rex L.

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AB - 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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A general purpose method for extracting RNA from Dictyostelium cells

Abstract

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ASJC Scopus subject areas

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