A reliable general purpose method for extracting genomic DNA from Dictyostelium cells.
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.
Pilcher
Karen E
KE
dictyBase, Center for Genetic Medicine, Northwestern University, 676 North Saint Clair Street Suite 1260, Chicago, Illinois 60611, USA.
Fey
Petra
P
Gaudet
Pascale
P
Kowal
Anthony S
AS
Chisholm
Rex L
RL
D000818:Animals; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D004023:Dictyostelium / Q000166:cytology* / Q000235:genetics*; D016678:Genome; D023281:Genomics / Q000379:methods*
D004247:DNA