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Micro 4140

Molecular Microbiology Laboratory

Credit Hour(s): 3 Units
Instructor(s): Bullwinkle
Offered: Autumn, Spring
Prerequisite(s): Micro 4100; Micro 4130 (can be taken concurrently)
Role in Microbiology Major: Core (Required)

Laboratory Topics:

  • Genetic exchange: transformation and conjugation
  • Plasmid vectors, cloning, and gel electrophoresis
  • Protein purification and SDS-PAGE
  • Immunoblotting
  • In vitro mutagenesis: Random and/or site-specific mutagenesis using PCR
  • In vivo mutagenesis: Transposon mutagenesis, recombineering
  • CRISPR/Cas tools
  • Gene reporters and in vitro enzyme assays
  • Bioinformatics


Learning Outcomes:

Successful students will be able to...

  • Handle and store microorganisms safely and maintain pure cultures using aseptic techniques
  • Describe and perform different mechanisms for genetic exchange in microbes
  • Apply basic calculations in the lab including dilutions, CFU counts, mutant frequency, in vitro reaction set up, master mixes, and unit conversion
  • Follow basic molecular biology protocols with technical accuracy
  • Design primer sequences and specify reaction parameters for PCR and similar protocols.
  • Describe and perform techniques for recombinant DNA construction, mutagenesis, and recombineering (PCR, restriction digestion, ligation, CRISPR)
  • Distinguish and design a genetic selection and a genetic screen
  • Interpret and manipulate DNA sequences to align, design primers, identify restriction enzyme recognition sites, and translate reading frames.
  • Recognize and compare mutation types in their data
  • Interpret gels and immunoblots, including the use of standards
  • Distinguish between positive and negative controls and recognize limitations in data and methods where experimental controls are needed.
  • Collect and analyze basic quantitative data via basic graphing programs
  • Communicate experimental and findings through writing and oral presentations
  • Apply concepts used in lab to interpreting other recent or advanced techniques in molecular microbiology