Team:Osaka/Protocols

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Protocols

  • Preparation of competent cells for heat-shock transformation
    • Preparation
  1. SOB medium 1L(-> final conc.)
    • Bacto Tryptone 20g(-> 2.0%)
    • Bacto Yeast Extract 5g(-> 0.5%)
    • 5M NaCl 2ml(-> 10mM)
    • 2M KCl 1.25ml(-> 2.5mM)
    • Add miliQ water 990ml to all miture and sterilize by autoclave. Before using, add sterilized Mg2 solution(1M MgSO4, MgCl2) 10 ml.
  • Transformation by heat-shock
  • Plasmid DNA miniprep
  • 3A BioBrick assembly
  • PCR cloning
  • PCR site-directed mutagenesis
  • Extraction of genome DNA for PCR cloning

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