Course Description
Laboratory methods stressing RNA and protein analysis techniques. Experiments and discussions will include RT-PCR, immunocytochemistry, flow cytometry, and other related experimental approaches. Topics of discussion will also include experimental design, pitfalls, limitations and advantages of experimental techniques, and data analysis and interpretation. Students will also learn to identify unknown samples utilizing learned approaches.
Athena Title
Exp Methods in Animal Biotech
Prerequisite
BCMB 3100 or BCMB 3100E or BCMB 3100H or MIBO 3500 or MIBO 3500E or MIBO 3500H or permission of department
Pre or Corequisite
BCMB 4010/6010
Semester Course Offered
Offered spring
Grading System
A - F (Traditional)
Course Objectives
The objective of this course is to give beginning graduate students hands-on training in basic cell culture, RNA, and protein laboratory techniques, including RT-PCR, immunocytochemistry, and flow cytometry. In this course, we will discuss the technical aspects of these approaches, experimental design, when to utilize techniques, limitations of these approaches, and other key components. Students will be given unknown cell derivatives to identify using newly learned skills to further enhance their understanding of the real-world applications of these techniques.
Topical Outline
Cell Culture 1. Lecture: Introduction to Cell Culture 2. Lab: The Essential Tools of Cell Culture 3. Lab: Maintaining and Passaging Cells from Primary Cells to Immortalized Cell Lines RNA Analysis 1. Lecture: Introduction to RT-PCR 2. Lab: Approaches to Successfully Isolating High Quality RNA 3. Lab: Reverse Transcription and Polymerase Chain Reaction 4. Lab: Gel Electrophoresis 5. Lab: Imaging and Analyzing Gels 6. Discussion: Student Presentation on Identified Unknown Samples Immunocytochemistry Protein Analysis 1. Lecture: Introduction to Immunocytochemistry 2. Lab: Fixing and Blocking 3. Lab: Antibody Incubations 4. Lab: Standard and Confocal Florescence Imaging 5. Lab: Imaging Analyzes 6. Discussion: Student Presentation on Identified Unknown Samples Flow Cytometry and Fluorescence Activated Cell Sorting Protein Analysis 1. Lecture: Introduction to Flow Cytometry and Fluorescence Activated Cell Sorting (FACS) 2. Lab: Fixed or Live Staining and Blocking 3. Lab: Antibody Incubations 4. Lab: Flow Cytometry 5. Lab: Analyzing Flow Cytometry Data 6. Lab: Cell Isolation Via FACS and Challenges of Post-FACS Culture 7. Discussion: Student Presentation on Identified Unknown Samples
Syllabus
Public CV