Many students either have minimal experience working in a laboratory or
none at all. This course introduces the prospective graduate student to general laboratory policies and basic procedures in order to excel in the biomedical sciences. Course Description This course will focus on providing students with a set of basic laboratory skills such as safety practices, laboratory mathematics, documentation, and good laboratory practices. Topics covered include solution chemistry; protein extraction and detection using Western blot and ELISA; the basic principles of immunohistochemistry; cell culture basics; and RNA extraction. Students will also be educated in research methodology, data analysis, and data presentation. This course is well suited to students in biomedical sciences. INTRODUCTION Learning Objectives: 1. Laboratory Security & Safety 2. Documentation: The Lab Notebook & Lab Report 3. Time management and multitasking 4. Learn the importance of writing recipes/protocols 5. Common lab equipment: calibration, accuracy and validation 6. Professionalism and teamwork 7. Laboratory organization Math Skills for the Laboratory Learning Objectives: 1. Introduction to the metric system: common units and prefixes 2. Learn to interpret positive and negative exponents 3. Review basic math skills that are commonly used in biomedical research 4. Learn to work with numbers in scientific notation 5. Introduction to Microsoft excel: Tips & Tricks Making Solutions and Dilutions Learning Objectives: 1. Recognize the importance of proper use of measuring instruments 2. Understand the concepts of mathematics of making solutions 3. Define various types of solutions: Buffers, reaction mixtures, cell culture media, cell lysates, liquid acids and bases 4. Correctly prepare a solution of a given molarity 5. Learn how to perform parallel and serial dilutions and how to determine dilution factor 6. Learn the concept of pH and buffers. 7. Learn how to adjust the pH of solutions.
Protein Detection I: Western Blot
Learning Objectives: 1. Gain general knowledge of different tissue/cell disruption methods 2. Learn how to determine total protein concentration in biological specimen 3. Become familiar with sample preparation 4. Be familiar with appropriate standards and controls 5. Learn how to prepare protein samples for Western Blot. 6. To become familiar with the basic theory of protein separation using polyacrylamide gel electrophoresis. 7. To become familiar with the basic theory behind protein membrane transfer. 8. Have a basic understanding of antibody usage: Immunodetection 9. Learn how to interpret results 10. Troubleshooting: Learn how to systematically and comprehensively approach problems 11. Learn how to present data
Protein Detection II: Enzyme Linked Immunosorbent Assay (ELISA)
Learning Objectives: 1. Have a basic understanding of antigen-antibody complex formation 2. To understand the steps involved in performing an ELISA 3. Gain general knowledge of different types of ELISA 4. Become familiar with the different detection methods including labeled enzymes and chromogenic substrates 5. Learn how to calculate results by developing standard curves 6. Learn how to measure the optical density of a solution using a spectrophotometer 7. How ELISA is used as a diagnostic tool Protein Detection III: Microscopy & protein immunostaining Learning Objectives: 1. Gain general knowledge with standard histological techniques including tissue fixation, processing and sectioning 2. Be familiar with the advantages and disadvantages of different fixation methods 3. Gain general knowledge of principles of immunohistochemical staining including immunofluorescence 4. Learn how to validate and optimize antibodies for various applications and various tissues. 5. Become familiar with the basic theory how antibodies maybe used to examine protein expression and distribution 6. Learn how to use the light microscope
7. Learn how to create figures for comparison of microscopic images and
illustration of important findings 8. Learn how to properly describe and interpret results of immunohistochemistry RNA Isolation Learning Objectives: 1. Understand the basic structure and function of RNA and its central role in gene expression 2. Appreciate the importance of RNA in biomedical research 3. Be familiar with Sample preparation for RNA isolation 4. Learn how to homogenize tissue: phenol and non-phenol based methods 5. Recognize that different tissue types require different homogenization methods 6. Precautions: Learn how to avoid sample RNase contamination 7. Understand the concepts behind quantitation of isolated RNA using UV spectroscopy 8. Learn how to asses RNA integrity and length by running RNA in a denaturing agarose gel 9. Acknowledge the importance of careful labeling and storage of RNA
Cell Culture Basics
Learning Objectives: 1. Introduction and overview of cell culture basics 2. Basic Equipment: laminar-flow hood and incubators 3. Gain general knowledge of aseptic techniques 4. Define primary cells 5. Learn how to prepare and sterilize media 6. Recognize different components of cell culture medium 7. Overview of appropriate cell line selection for experiments 8. Learn how to maintain and cryopreserve cell Lines 9. Recognize and troubleshoot problems: Contamination