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Abstract: A rapid "miniprep" method allowing a simultaneous analysis of DNA RFLP markers and dsRNA molecules was perfected in a wild-type strain of the cultivated basidiomycete A. aegerita. Biological material consisted of dikaryotic mycelium taken from a single Petri dish. Cell wall disruption was carried out by cmshing the mycelium in liquid nitrogen in a mortar and pestle. Extraction was realized by using CTAB as detergent. CTAB was removed after ethanol precipitation by three washings with 70% ethanol. Hence, total nucleic acid extracts were obtained without phenol treatment. In routine work, twelve strains are tested a day. 0.2 g of fresh mycelium furnishes 1 mg of total nucleic acids (Nuclear and mitochondrial DNA, RNA and dsRNAs).The dsRNA molecules were visualised by electrophoresis of the extract after its treatment by DNase I and RNase A in high salt concentration (300 mM NaCl). The method was suitable to characterize dikaryotic, homokaryotic strains as well as basidiocarps, by RFLP markers and to detect simultaneously the presence of dsRNAs in these different types of mycelium.
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