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DNA

General techniques

  • Procedure for incoming plasmids
  • Preparation of glycerol stocks
  • Ethanol precipitation of DNA or RNA
  • Phenol-Chloroform extraction of nucleic acids
  • Cleavage of DNA into mononucleotides

Plasmid DNA isolation

  • Minipreps using Promega kit (also available in french)
  • Midipreps using Macherey & Nagel Kit
  • BAC miniprep

Plasmid manipulation

  • Ligation (also available in french): excel sheet for calculations
  • Gibson assembly: excel sheet for calculations
  • Annealing and ligation of linkers
  • Recombination of BAC with PCR product
  • Restriction enzyme digestion (also in french with agarose gel elecrophoresis)
  • Dephosphorylation of plasmids

Transformation and preparation of competent cells

  • Preparation of electrocompetent cells
  • Electroporation of electrocompetent cells
  • Rapid preparation of electrocompetent cells
  • Preparation of chemically competent cells  (also in french)
  • Transformation into chemically competent cells

Extraction of DNA from mammalian cells:

  • genomic DNA extraction
    • 12-well plate
    • 96-well plate
    • mouse tail
    • Isolation of DNA from TRIZOL
  • extrachromosomal DNA
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Guido Bommer, de Duve Institute, Université Catholique de Louvain, Woluwe St. Lambert, Belgium