A general purpose method for extracting RNA from Dictyostelium cells.
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.
Pilcher
Karen E
KE
dictyBase, Center for Genetic Medicine, Northwestern University, 676 North Saint Clair Street Suite 1260, Chicago, Illinois 60611, USA.
Gaudet
Pascale
P
Fey
Petra
P
Kowal
Anthony S
AS
Chisholm
Rex L
RL
D000818:Animals; D002725:Chloroform / Q000737:chemistry; D004023:Dictyostelium / Q000737:chemistry / Q000235:genetics*; D019800:Phenol / Q000737:chemistry; D012313:RNA / Q000737:chemistry / Q000302:isolation & purification*; D013997:Time Factors
D019800:Phenol; D012313:RNA; D002725:Chloroform