Protocol abstract
Nature Protocols 2, - 1329 - 1332 (2007)
Published online: 24 May 2007 | doi:10.1038/nprot.2007.191
Subject Categories: Isolation, purification and separation | Model organisms | Nucleic acid based molecular biology
A general purpose method for extracting RNA from Dictyostelium cells
Karen E Pilcher1, Pascale Gaudet1, Petra Fey1, Anthony S Kowal2 & Rex L Chisholm1,2
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.
- dictyBase, Center for Genetic Medicine, Northwestern University, 676 North Saint Clair Street Suite 1260, Chicago, Illinois 60611, USA.
- Center for Genetic Medicine, Northwestern University, 303 East Superior Room 7-125, Chicago, Illinois 60611, USA.
Correspondence to: Rex L Chisholm1,2 e-mail: r-chisholm@northwestern.edu
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