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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
