M4140

Molecular Microbiology Laboratory


Credit: 3 units

Instructor: Bullwinkle

Offered: Autumn, Spring

Laboratory Topics:

  • Antibody purification and antibody reactions
  • Enzyme-linked immunosorbent assay (ELISA)
  • Immunofluorescense microscopy and fluorescence-activated cell sorting (FACS)
  • Immunoblotting
  • Genetic exchange: transformation and conjugation
  • In vitro mutagenesis: Random and site-specific mutagenesis using PCR
  • In vivo mutagenesis: transposon mutagenesis
  • Phenotypic and molecular analysis of transposon mutants
  • Bioinformatics

Learning Outcomes:

Successful students will be able to...

  • Handle microorganisms safely and maintain pure cultures using aseptic techniques (aseptic transfers, three phase streak)
  • Manage and preserve a collection of microorganism strains
  • Describe and perform the three mechanisms for genetic exchange in microbes (transformation, transduction, conjugation)
  • Distinguish and design a genetic selection and a genetic screen
  • Calculate mutation frequencies
  • Extract DNA (genomic, plasmid) from bacteria using commercial kits
  • Clone and/or sequence the DNA adjacent to the site of a transposon insertion
  • Amplify DNA by PCR, including designing the sequences of primers and specifying the parameters of the amplification reaction
  • Interpret and manipulate DNA sequence data in silico to identify restriction enzyme recognition sites, translate DNA to protein sequence, align both DNA and protein sequences, and BLAST DNA sequence to identify genomic fragments
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