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# Course Rationale

## 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
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
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