A simple method for isolation of nuclei from <Emphasis Type="Italic">Basidiobolus ranarum</Emphasis> (Zygomycota) |
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| Authors: | Mio Nakamura Koji Nomura Jun-ichi P Abe Yousuke Degawa Makoto Kakishima |
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| Institution: | 1.Graduate School of Life and Environmental Sciences,University of Tsukuba,Tennodai, Tsukuba, Ibaraki,Japan |
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| Abstract: | A simple method for isolating the nuclei from Basidiobolus ranarum was established. To improve the yield and purity of nuclei, we investigated maceration methods, buffer composition, and centrifugation
conditions to establish an optimal procedure. Basidiobolus ranarum cultured for 5 days was enzymatically macerated and then homogenized and filtrated through stainless steel sieves. The crude
cell homogenate was loaded on a layer of buffer containing 50% glycerol and centrifuged at 1500 g. The resultant pellet contained pure nuclei. |
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| Keywords: | Cell fractionation Glycerol density-gradient centrifugation Nucleus |
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