Laboratory Manual for Biotechnology and Laboratory Science The Basics

The authors of this manual are faculty in the Biotechnology Laboratory Technician Program at Madison Area Technical College, which prepares students to become laboratory professionals. These authors have years of experience and have spent a great deal of time in the classroom, trying out the exercises and activities which are laid out in the manual. Lisa Seidman is highly respected for her work in biotechnology instruction.

I. SAFETY IN THE LABORATORY

Unit Introduction

SAFETY PART 1: CREATING A SAFE WORKPLACE     

Fundamental Principles                                                                                     

Classroom Activity 1: Performing a Risk Assessment                                          

Classroom Activity 2: Exploring the CDC and OSHA Websites                                         

Classroom Activity 3: Responding to Emergencies                                                                                                                                   

SAFETY PART 2: WORKING SAFELY WITH CHEMICALS

Fundamental Principles

Classroom Activity 4: Understanding the Chemicals with which You Work 

Classroom Activity 5: Personal Protection                                                                       

Laboratory Exercise 1: Tracking the Spread of Chemical Contamination                 

Classroom Activity 6: Analyzing Safety Issues in a Laboratory Procedure                                                  

SAFETY PART 3: WORKING SAFELY WITH BIOLOGICAL HAZARDS

Fundamental Principles                                                                                     

Laboratory Exercise 2: Production of Bioaerosols and Factors Affecting Aerosol Production

Unit Discussion                                                                                                

Safety Contract, student copy

Safety Contract, to turn in

II. DOCUMENTATION IN THE LABORATORY      

Unit Introduction                                                                                                

Classroom Activity 7: Be an Auditor                                                                    

Classroom Activity 8: Writing and Following an SOP                                            

Laboratory Exercise 3: Keeping a Laboratory Notebook                                                    

Unit Discussion

III. METROLOGY IN THE LABORATORY            

Unit Introduction                                                                                                                                                                                                                                    

Laboratory Exercise 4: Recording Measurements with the Correct Number of Significant Figures                                                                                                                 

Classroom Activity 9: Constructing a Simple Balance                                           

Laboratory Exercise 5:Weight Measurements 1; Good Weighing Practices           

Laboratory Exercise 6: Weight Measurements 2; Performance Verification

Laboratory Exercise 7: Volume Measurements 1; Proper Use of Volume Measuring Devices

Laboratory Exercise 8: Volume Measurements 2; Performance Verification of a Micropipette

Laboratory Exercise 9: Measuring pH with Accuracy and Precision                                   

Unit Discussion                                                                                                                                                                                                                                    

IV. SPECTROPHOTOMETRY                

Unit Introduction                                                                                                

Laboratory Exercise 10: Color and the Absorbance of Light                                   

Laboratory Exercise 11: Concentration, Absorbance, and Transmittance    

Laboratory Exercise 12: Preparing a Standard Curve with Food Coloring and Using it for Quantitation           

Classroom Activity 10:Beer’s Law and Calculating an Absorptivity Constant                                  

Laboratory Exercise 13: Determination of the Absorptivity Constant for ONP                                  

Unit Discussion                                                                                                                                                                                                                                                

V. BIOLOGICAL SOLUTIONS                

Unit Introduction                                                                                                

Classroom Activity 11: Getting Ready to Prepare Solutions with One Solute: Calculations Classroom Activity 12: Getting Ready to Prepare Solutions with One Solute: Ordering Chemicals           

Laboratory Exercise 14: Preparing Solutions with One Solute

Laboratory Exercise 15: Preparing Solutions to the Correct Concentration  

Laboratory Exercise 16: Working with Buffers                                                      

Laboratory Exercise 17: Preparing Breaking Buffer                                              

Laboratory Exercise 18: Preparing TE Buffer                                                        

Laboratory Exercise 19: More Practice Making a Buffer                

Laboratory Exercise 20: Making a Quality Product in a Simulated Company                      

Unit Discussion                                                                                                                                                                                                                                                

VI. ASSAYS                 

Unit Introduction                                                                                                

Laboratory Exercise 21: Two Qualitative Assays                                      

Laboratory Exercise 22:UV Spectrophotometry of DNA; Quantitative Applications

Laboratory Exercise 23:UV Spectrophotometry of DNA, RNA, and Proteins; Qualitative Applications

Laboratory Exercise 24: Using Spectrophotometry for Quality Control; Niacin

Laboratory Exercise 25: A Quantitative Protein Assay; Learning the Assay

Laboratory Exercise 26: Exploring Assay Verification                                           

Laboratory Exercise 27: The Beta-Galactosidase Enzyme Assay              

Laboratory Exercise 28: Comparing the Specific Activity of Two Preparations of Beta-Galactosidase

Unit Discussion                                    

VII. SEPARATION METHODS                

Unit Introduction                                                                                                

Classroom Activity 13: Separating Materials Using a Centrifuge, Planning

Laboratory Exercise 29: Separation of Two Substances Based on Their Differential Affinities for Two Phases                                                                                                

Laboratory Exercise 30: Separation of Dyes Using Paper Chromatography

Laboratory Exercise 31: Separating Molecules by Agarose Gel Electrophoresis

Laboratory Exercise 32: Using Agarose Gel Electrophoresis to Perform an Assay

Laboratory Exercise 33: Optimizing Agarose Gel Electrophoresis

Laboratory Exercise 34: Quantification of DNA by Agarose Gel Electrophoresis

Unit Discussion

VIII. GROWING CELLS 

Unit Introduction

WORKING WITH BACTERIAL CELLS

Laboratory Exercise 35: Aseptic Technique on an Open Lab Bench 

Laboratory Exercise 36: Working with Bacteria on an Agar Substrate; Isolating Individual Colonies

Laboratory Exercise 37: Using a Compound Light Microscope

Laboratory Exercise 38: Gram Staining

Laboratory Exercise 39: Making Phosphate Buffered Saline

Laboratory Exercise 40: The Aerobic Spread Plate Method of Enumerating Colony Forming Units

Laboratory Exercise 41: Generation of an E. coli Growth Curve

WORKING WITH MAMMALIAN CELLS

Laboratory Exercise 42: Aseptic Technique in a Laminar Flow Hood

Laboratory Exercise 43: Making Ham’s F12 Medium from Dehydrated Powder

Laboratory Exercise 44: Examining, Photographing, and Feeding CHO (Chinese Hamster Ovary) Cells

Laboratory Exercise 45: Counting Cells Using a Hemacytometer

Laboratory Exercise 46: Subculturing CHO Cells

Laboratory Exercise 47: Preparing a Growth Curve For CHO Cells

Unit Discussion

APPENDICES

I Materials and Supplies that are Assumed to be Present in Your Laboratory

II Calculating Standard Deviation                                                                                     

II Periodic Table of Elements

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