A method for isolation of cytoplasmic RNA from a slime mold,Physarum polycephalum |
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| Affiliation: | 1. Department of Civil and Environmental Engineering, University of California, Los Angeles, CA 90095, United States;2. Department of Civil and Environmental Engineering, University of Pittsburgh, Pittsburgh, PA 15260, United States;3. Department of Biological Chemistry, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, United States;4. California NanoSystems Institute, University of California, Los Angeles, CA 90095, United States |
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| Abstract: | A procedure for fast and simple preparation of cytoplasmic ribonucleic acid from Physarum polycephalum microplasmodia is described. Microplasmodia are homogenized in a high-magnesium-high-ethylene glycol bis(β-aminoethyl ether) N,N′-tetraacetic acid buffer and nuclei are pelleted. The supernatant is extracted with sodium dodecyl sulfate-phenol-chloroform and crude RNA is precipitated. This is further purified by selective ethanol precipitation from 6 m guanidinum hydrochloride. This RNA preparation is suitable for direct use in hybridization studies. |
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