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Methodology and Technical Support

Bacillus subtilis - detekce mutátorů (metody)

Methodology

  • Conventional microbiological methods are used for the handling of the bacterial cultures (laboratory strains, soil and clinical isolates), providing physiologically precisely defined cultivations.
  • Molecular genetic methods such as specific and non-specific mutagenesis, construction of genomic libraries, etc., are used to search for novel genes.
  • Genetic engineering methods are used to generate fusion with reporter genes (beta-galactosidase, GFP) for monitoring promoter activities in vivo, cloning, PCR, Southern and Northern blotting.
  • We also use SDS-PAGE (1D and 2D), Western blotting with immunodetection and qPCR for monitoring levels of gene expression, and flow cytometry for monitoring gene expression in subpopulations.
  • Light, fluorescence and electron microscopy are also essential techniques in microbiology.
  • A bioinformatics analysis of the genome dynamics of different strains of Bacillus and Stenotrophomonas, as well as an analysis of evolutionary relationships of selected genetic determinants are used to examine various relationships.

 

Technical Support

  • The devices for both small and large-scale cultivation of bacteria
  • Thermocycler
  • Horizontal agarose gel electrophoretic systems
  • 1D SDS-PAGE (+ detection systems)
  • Southern, Northern, Western blotting systems
  • Light microscope, laminar box, thermostats, centrifuges, deep-freeze box etc.
  • The technical equippment of the Biological Section is used for other, more sophisticated methods.

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