• Title/Summary/Keyword: Alkaline Iysis

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Improved Procedure for Large-scale Isolation of Mitochondrial DNA from Mammalian Tissues

  • Hong, Sung-Soo;Lee, Chung-Choo
    • Animal cells and systems
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    • v.3 no.1
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    • pp.73-78
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    • 1999
  • Although there are several methods for the preparation of mitochondrial DNA (mtDNA) from mammalian tissues, most are relatively long ultracentrifugation or manipulations by a small-scale method. We escribed a rapid method for large-scale extraction of mtDNA from human placental and horse liver tissues. The method is based on the preparation and homogenization of tissues, urification of crude mitochondria by differential centrifugations and isolation of mtDNA by alkaline Iysis. It was improved from Pre-existing methods by replacing some steps with simpler ones and discarding many others. This method gives a high yield of pure mtDNA(approximately 1-5mg from one placenta; ca. 400-600 g wet weight), depending on its sources (fresh tissue gave better results than frozen one). The resulting mtDNA indicated that this method can yield mtDNA in sufficient purity and quantity to identify the direct restriction analysis on agarose gel, random-primed labeling as a probe, and end labeling. Therefore, the method is ideal for obtaining good mtDNA samples to conduct routine restriction fragment length polymorphism (RFLP) analyses of natural populations for genetic studies.

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Incidence of Plasmids in Marine Bacteria Isolated from the Bunker-C Oil Enriched Culture (Bunker-C유 집식배양으로부터 해양세균 Plasmid의 분포)

  • Park, In-Sick;Park, Jung-Youn;Jin, Deuk-Hee;Hong, Yong-Ki
    • Microbiology and Biotechnology Letters
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    • v.16 no.4
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    • pp.270-274
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    • 1988
  • Samples used for the enrichment culture were collected from the sea water of suspected chronic petroleum contamination in the vicinity of Pusan, Chungmu and Ulsan ports, Korea. Alkaline Iysis and agarose gel electrophoresis techniques were employed to screen these isolates for the presence of plasmid DNA. There were n little differences in the percentage of isolates containing plasmids between sampling sites of unpolluted sen water (22%) and polluted son water (25%). Bacterial isolates taken from the Bunker-C oil enriched culture showed significantly more plasmid incidence (29%). About two thirds of strains grown on a variety of hydrocarbons were Gram negative strains of which 33% contained one or more plasmids. Multiple plasmids were observed in 23% of the plasmid-carrying strains. Forty one percent of the plasmids detected were estimated to have a mass of 20 kb or more.

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