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An easy and rapid method for isolation of entire mycobacterial genome for application in pulsed-field gel electrophoresis

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Abstract

A DNA extraction suitable for mycobacterial lysis in gentle conditions compatible with genome analysis by pulsed-field gel electrophoresis is presented. Effects of preliminary treatments with SDS, Triton X-100, and hexane on mycobacterial outer layer were observed by electron microscopy. The most efficient procedure, performed on cells from liquid or solid medium, consisted of treatment by Triton X-100, agarose embedding of the cells, and further treatment with α-amylase followed by lysozyme and SDS-proteinase K.

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Authors and Affiliations

  1. Unité de la Tuberculose et des Mycobactéries, Institut Pasteur, 75724, Paris Cedex 15, France

    Martine Charvin, Nalin Rastogi &  Véronique Vincent Lévy-Frébault

Authors
  1. Martine Charvin
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  2. Nalin Rastogi
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  3. Véronique Vincent Lévy-Frébault
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Charvin, M., Rastogi, N. & Lévy-Frébault, V.V. An easy and rapid method for isolation of entire mycobacterial genome for application in pulsed-field gel electrophoresis. Current Microbiology 22, 327–331 (1991). https://doi.org/10.1007/BF02091963

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  • DOI: https://doi.org/10.1007/BF02091963

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