PHC 542 (1 credit)
Contemporary Methods in the Pharmaceutical Sciences:
3 - Quantitative RNA Approaches

Course Coordinator:
            Kathleen M.K. Boje, Ph.D.; Assistant Professor of Pharmaceutics

Lecturers:
            Kathleen M.K. Boje, Ph.D.; Assistant Professor of Pharmaceutics
            Debra Dubois, Ph.D.; Research Assistant Professor of Biology
            Richard R. Almon, Ph.D.; Associate Professor of Biology

There are four major topics for the course:
1. RNA: Overview and Isolation / Enrichment Techniques
2. Qualitative RNA Analysis Methods
3. Quantitative RNA Analysis Methods
4. Application of Quantitative RNA Analysis to Pharmacokinetic/ Pharmacodynamic Studies of Gene Expression

1. RNA: Overview and Isolation / Enrichment Techniques

Class 1. Why Study RNA? An Overview
Classes 2 & 3: Planning an RNA Extraction Experiment; Strategies for RNA Enrichment

Mini-Laboratory Exercises
Lab 1. Isolation of Cytoplasmic RNA with NP-40 from Cultured Cells
Lab 2. Isolation of Cellular RNA with GTC-Acid-Phenol from Cultured Cells

2. Qualitative RNA Analysis Methods
Class 4: Isolation of Total RNA from Tissue
Class 5: Isolation of Poly(A+)RNA by Various Technologies
Classes 6: Spectrophotometric Analysis of Nucleic Acids and Qualitative Northerns
Classes 7 & 8: Quantitative Northern Blots

Mini- Laboratory Exercise
Lab 3. Determination of RNA in NP-40 and GTC-Acid-Phenol Preps; Qualitative Northern Analysis I: RNA Electrophoresis or NP-40 or GTC-Acid-Phenol Preps;
Lab 4: Northern Analysis II: Inspection of Gels; Blotting
Lab 5. Northern Analysis III: Prehybridization; Hybridization and Chemiluminescent Detection

3. Quantitative RNA Analysis Methods
        Classes 9 & 10: Quantiative Northern Blots

4. Application of Quantitative RNA Analysis to Pharmacokinetic/ Pharmacodynamic Studies of Gene Expression

Classes 12 & 13: Quantitative RNA Analysis in Pharmacokinetic /Pharmacodynamic Studies

Course Evaluation: This course will be letter graded by the criteria set forth by the University of Buffalo. A
course grade will be determined by the student's performance on announced quizzes given after each topic.

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