This course will begin with interactive training modules in which you will be introduced to the methodologies currently used in biomedical research laboratories, including (but not limited to) basics of molecular biology and protein science, genetic analysis, cell culture, microscopy, mutagenesis, phenotypic analysis, immunoassays and protein purification. In order to understand how these techniques are used to solve research problems, you will be applying these methodologies to unique training projects developed specifically for this course.

Upon completion of the training modules, you will then have the invaluable opportunity to directly apply the techniques and analytical skills you acquired in the training lab to two independent scientific projects, each of which that will focus on a distinct research area under the supervision of a professor from the Faculty of Medicine.

By the end of this laboratory course, the student will be able to:

• Acquire experience in common laboratory techniques routinely used in biomedical research labs
• Define experimental hypotheses to guide inquiry-based projects and assess the primary literature to gauge the current state of diverse disciplines
• Design controlled assays to address specific experimental questions and predict expected outcomes
• Interpret primary data acquired from a range of experimental techniques and evaluate quality of research data produced by peers and the scientific community
• Clearly communicate scientific data in written and verbal forms and actively participate in scientific discussion
• Collaborate with peers on group-based projects and contribute to the laboratory learning environment
• Apply safe laboratory methods to ensure the safety of self, coworkers and the environment

TMM3009 is divided in two sections:

Part I: Experimental Training

In this section, students will develop technical and analytical skills in a teaching laboratory environment. This section of the course consists of four modules, each of which focuses on a unique research project.

Part II: Rotations

In this section, students will have the opportunity to conduct research projects in two different labs in the Faculty of Medicine, each of which focuses on distinct biomedical processes.

Module 1: Introduction to biomedical laboratory techniques

Objective: By the end of this module, the student will be able to perform fundamental techniques in molecular and cellular biology commonly used in the biomedical laboratory.

Experimental techniques: Pipetting, bacterial transformations, plasmid isolation, PCR, restriction digestion, agarose gel electrophoresis, mammalian tissue culture, transfections, SDS-PAGE, Western blotting, immunofluorescence, microscopy and image acquisition.

Theoretical concepts: Module 1 is divided into two sections. The first section (Weeks 1 &2) will introduce students to a range of basic lab techniques in molecular biology, cell culture, microscopy, and protein analysis. The second section (Weeks 3 & 4) will allow students to apply the techniques introduced in the first two weeks to address an experimental question. 

Module 2: Testing if phosphorylation of the Sir3 heterochromatin protein modulates the disassembly of repressive chromatin.

Objective: The goal of this module is to test if known and putative phosphorylation sites on Sir3, a yeast heterochromatin protein, modulate the disassembly of SIR-dependent silent chromatin. We will examine phosphosite mutants using a sensitive fluorescent reporter that allows the resolution of individual de-repression events.

Experimental techniques: Oligonucleotide design, PCR, yeast husbandry, Cas9-mediated recombination, western blotting, chromatin immunoprecipitation, quantitative PCR, yeast growth assays, live microscopy.

Theoretical concepts: Mutagenesis, CRISPR-Cas9, basic yeast cell biology and genetics, epigenetic regulation of chromatin structure, antibodies, counter-selection.

Module 3 - Purification and analysis of a metalloregulator

Objective: The goal of this module is to introduce basic concepts of protein biology and to understand how protein structure can impact protein function.

Experimental techniques: Mutagenesis, electrophoretic mobility shift assays, affinity purification, protein gels and coomassie staining.

Theoretical concepts: Fundamental concepts in protein biology, protein structure and functional analysis and protein binding kinetics.

Following the experimental training portion of the course, the student will complete two rotation projects under the supervision of faculty members from the BMI or CMM departments at the Faculty of Medicine. Here, the student will have the invaluable opportunity to apply the experimental and scientific communication skills that they have learned in the teaching lab to conduct their own research projects. Each student will complete their rotations in two distinct laboratories that focus on different areas of biomedical research.