A method for the preparation of high-molecular-weight DNA from marine and freshwater triclads (Platyhelminthes, Turbellaria).
Although the last decade has seen much activity devoted to the phylogeny of the Platyhelminthes, such activity has relied in the main on traditional anatomical, ultrastructural, and developmental data. Extension of these studies to the molecular level has been impeded by the lack of a reliable method for the isolation of DNA from these organisms, especially marine triclads whose DNA is particularly difficult to isolate using methods currently used with other eukaryotic or prokaryotic organisms. We report here the details of a method that results in the rapid isolation of relatively pure DNA in good yields from small amounts of material. The method involves the treatment of the animals with high concentrations of NaDodSO4 followed by phenol extraction, resulting in a product that is an excellent substrate for restriction endonuclease digestion and Southern blotting.
Hempstead
P G
PG
Department of Biology, Memorial University of Newfoundland, St. John's, Canada.
Regular
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Ball
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D000818:Animals; D015139:Blotting, Southern; D004247:DNA / Q000302:isolation & purification*; D008970:Molecular Weight; D010932:Planarians / Q000235:genetics*; D012150:Polymorphism, Restriction Fragment Length; D014419:Turbellaria / Q000235:genetics*
D004247:DNA