A Laboratory Skills Course, Second Edition Detailed Chapter Outlines

Biotechnology: A Laboratory Skills Course, Second Edition Detailed Chapter Outlines

Below is a chapter-by-chapter outline of the student edition of Biotechnology: A Laboratory Skills Course, second edition. The vignettes and student activities are listed at the end of each chapter outline. Please visit bio-rad.com/textbook for more information.

Chapter 1: Biotechnology Industry 1.5 Industry Practices ■■ Good Laboratory Practice 1.1 What Is Biotechnology? ■■ Good Manufacturing Practice ■■ The Biotechnology Toolkit 1.6 Biotechnology Company Structure 1.2 Who Uses Biotechnology? ■■ Life Science Research 1.7 Careers in Biotechnology — -Omics and Systems Biology ■■ Careers in the Laboratory — Laboratory Technician ■■ Healthcare — Quality Assurance Technician — Drug Discovery, Development, and Testing — Research Associate — Protein-Based Drug Production — Research and Development Scientist — Personalized Medicine — Clinical Scientist — Clinical Diagnostics — Engineer ■■ Agriculture ■■ Non-Laboratory Career ■■ Food ■■ Industrial Manufacturing Chapter 1 Vignettes ■■ Biofuels Bioethics — Biotechnology: Good or Bad? ■■ Mining Biotech in the Real World — Affecting Government Policy ■■ Pollution Monitoring and Waste Management ■■ Conservation Biotech in the Real World — Bio-Rad: Then and Now ■■ Biodefense Careers in Biotech — Timothy Balmorez, Chemist, Bio-Rad ■■ Forensics Laboratories, Inc. ■■ Human Origins ■■ Nanotechnology How To… Write an SOP 1.3 The Biotechnology Industry ■■ What Is a Biotechnology Company? Chapter 2: Laboratory Skills

■■ Biotechnology Product Development 2.1 Laboratory Safety ■■ Funding Biotechnological Advancement ■■ Operating Safely in a Laboratory ■■ Patents ■■ Safety Training 1.4 Governmental Regulation of Biotechnology ■■ Personal Protective Equipment ■■ Regulation of Genetically Modified Organisms ■■ Chemical Safety ■■ Regulation of Products in Healthcare ■■ Biological Safety ■■ Preclinical Research ■■ General Laboratory Safety ■■ Clinical Trials ■■ Waste Disposal ■■ Regulation of Animal Research ■■ Cleanrooms

■■ Regulation of Human Research 2.2 Laboratory Notebooks ■■ Regulation of Waste Disposal ■■ Laboratory Notebook Structure ■■ Regulation of Nanotechnology ■■ Components of a Laboratory Notebook Entry ■■ Workplace Safety Regulation Biotechnology: A Laboratory Skills Course, Second Edition

2.3 Laboratory Equipment Chapter 2 Vignettes ■■ Measuring volumes Bioethics — Waste Disposal — Graduated Cylinders Biotech in the Real World — Who Invented Claritin? How — Volumetric Flasks Laboratory Notebooks Played Their Part — Vacuum-Assisted Pipetts — Micropipets Careers in Biotech — Joshua Moore, Bio-Containment — Burettes Protocol Support Supervisor, U.S. Army Medical Research Institute of Infectious Diseases (USAMRIID) ■■ Liquid Containers — Erlenmeyer Flasks How To… Use an Adjustable-Volume Micropipet — Reagent Bottles Chapter 2 Activities ■■ Measuring Mass 2.A DNA Extraction and Precipitation — Balances 2.B Pipetting ■■ Measuring Temperature — Thermometers 2.C Kool-Aid Column Chromatography

■■ Measuring pH 2.D Making Solutions — pH Meters 2.E Titration ■■ Calibration 2.F Writing a Standard Operating Procedure ■■ Washing Glassware ■■ Disinfecting and Sterilizing — Autoclaving Chapter 3: Microbiology and Cell Culture — Disinfecting Surfaces with Bleach 3.1 Microbiology and Cell Biology — Disinfecting Surfaces with Alcohol ■■ Three Domains of Life — Disinfecting Using a Microwave Oven ■■ Microorganisms and History — Disinfecting with Germicidal UV Light — Sterilizing Solutions 3.2 Bacteria ■■ Names and Shapes of Bacteria ■■ Labeling ■■ Bacterial Environments 2.4 Numerical Data 3.3 Uses of Bacteria in Biotechnology ■■ Significant Figures ■■ Food Production ■■ Scientific Notation ■■ Protein Production ■■ Units of Measure — Metric Prefixes 3.4 Culturing Bacteria in the Laboratory ■■ Growth Media 2.5 Preparing Solutions — Solid Media ■■ Percent Solutions — Antibiotic Selection — Mass per Volume Percent Solutions — Liquid Media — Volume per Volume Percent Solutions — Percentages, Ratios, and Proportions 3.5 Microbiological Techniques — Diluting Percent Solutions ■■ Tools ■■ Aseptic Technique ■■ Using Stock Solutions Given in Terms of “x” ■■ Using Streak Plate Technique to Isolate ■■ Molar Solutions — The Mole Single Colonies ■■ — Molarity Labeling Media ■■ — Making Molar Solutions Quantifying Bacteria a. Proportional Method — Serial Dilution and Plate Counts b. Unit Cancellation Method — Optical Density of Bacteria ■■ Identification of Bacteria ■■ Making Compound Molar Solutions ■■ Diluting Molar Solutions 3.6 Eukaryotic Cells ■■ Preparing Small Volume Dilutions ■■ Organelles ■■ Normality ■■ Other Eukaryotic Cells

3.7 Uses of Eukaryotic Cells in Biotechnology 4.4 DNA Analysis Techniques ■■ Fermentation ■■ Horizontal Agarose Gel Electrophoresis ■■ Protein Production ■■ Agarose Gels and Running Buffer ■■ Stem Cells ■■ Standards for DNA Electrophoresis ■■ Equipment 3.8 Eukaryotic Cell Culture — Horizontal Gel Electrophoresis Chambers ■■ Growth Media — Power Supplies ■■ Environment ■■ Sterility ■■ Running an Agarose Gel ■■ Visualization ■■ Agarose Gel Staining and Imaging Chapter 3 Vignettes 4.5 Restriction Enzymes as Forensic Tools Bioethics — Should Human Embryos Be Used for Research? ■■ Restriction Fragment Length Polymorphisms ■■ Southern Blotting Biotech in the Real World — Artificial Life! Chapter 4 Vignettes Careers in Biotech — Katie Dalpozzo, Laboratory Research Bioethics — Personal Genetic Information Associate, Bavarian Nordic Biotech in the Real World — Fighting Crime with DNA How To… Use Aseptic Technique to Transfer Bacteria Careers in Biotech — Elisa Ciullo, Sr. Supervisor, Clinical Chapter 3 Activities Filing Operations, Genentech 3.A Making Microbiology Media How To... Set Up a Restriction Digest 3.B Disk Diffusion Test (Modified Kirby-Bauer Test) Chapter 4 Activities 3.C Microbes and Health: An Illustration of Koch’s Postulates 4.A Restriction Site Prediction Using NEBcutter 3.D Gram Staining 4.B Casting Agarose Gels 3.E Quantifying Bacterial Numbers 4.C Dye Electrophoresis 3.F Staining Eukaryotic Cells 4.D Restriction Digestion and Analysis of Lambda DNA

Chapter 4: DNA Structure and Analysis 4.E Forensic DNA Fingerprinting 4.F Plasmid Mapping 4.1 Molecular Biology ■■ The Central Dogma of Molecular Biology Chapter 5: Bacterial Transformation 4.2 DNA Structure and Plasmid Purification 4.3 Recombinant DNA Technology 5.1 History of Bacterial Transformation and ■■ Restriction Enzymes Plasmids Bacteria ■■ Ligases ■■ Plasmid Structure ■■ Nontraditional Cloning Techniques ■■ Features of Plasmids — Golden Gate Assembly ■■ Using Plasmids in Biotechnology — Gibson Assembly ■■ Transcriptional Regulation of Plasmids — TOPO (TA) Cloning ■■ Genetically Engineering the pGLO Plasmid ■■ Using Enzymes in the Laboratory ■■ Plasmids for Eukaryotic Expression ■■ CRISPR-Cas9 Technology — How CRISPR works 5.2 Transforming Cells ■■ — Applications of CRISPR Technology Selection of Transformed Cells ■■ a. Agriculture Transformation Efficiency b. Industry 5.3 Plasmid Purification and Quantitation c. Public Health ■■ Growing Bacteria in a Liquid Culture d. Medicine ■■ Purifying Plasmid DNA from a Culture ■■ DNA Quantitation — Gel Quantitation — Spectrophotometric Quantitation — Quantitation of DNA with a Fluorometer

Chapter 5 Vignettes 6.6 Techniques Based on PCR Bioethics — A World without Antibiotics? ■■ DNA Microarrays ■■ DNA Sequencing Biotech in the Real World — Biotech on the Pharm — Primer Sequences Careers in Biotech — Denise Gangadharan, PhD, Associate ■■ Next-Generation Sequencing Director for Science, Centers for Disease Control and — Sequencing by Synthesis Prevention — Pyrosequencing How To... Transform E. coli Using Electroporation — Ion Semiconductor Sequencing — Sequencing by Ligation How To… Use a Microcentrifuge ■■ Sequence Data Analysis Using Bioinformatics Chapter 5 Activities 5.A Bacterial Transformation with S3 Plasmid 6.7 Real-World Applications of PCR ■■ PCR in Medicine 5.B Bacterial Transformation with pGLO Plasmid ■■ PCR in Agriculture 5.C Purification of S3 and pGLO Plasmids ■■ PCR in Forensics ■■ PCR in Paternity Testing 5.D DNA Quantitation ■■ PCR in Human Migration ■■ PCR in Wildlife Conservation Chapter 6: The Polymerase Chain Reaction Chapter 6 Vignettes 6.1 Invention of PCR Bioethics — Forensic DNA Databases — Is Your Privacy Protected? 6.2 What Is PCR? ■■ Three Stages of PCR Biotech in the Real World — A Universe of Viruses — Denaturation Careers in Biotech — Dora Barbosa, Research Associate, — Annealing University of California San Francisco — Extension How To... Program a Thermal Cycler ■■ PCR and DNA Replication ■■ Components of a PCR Reaction Chapter 6 Activities ■■ Setting Up a PCR Reaction 6.A STR PCR Analysis — Master Mixes 6.B GMO Detection by PCR ■■ Analysis of PCR Products 6.C Detection of the Human PV92 Alu Insertion 6.3 Thermal Cyclers 6.D Fish DNA Barcoding 6.4 Types of PCR ■■ Real-Time, or Quantitative, PCR Chapter 7: Protein Structure and Analysis ■■ Digital PCR ■■ Reverse Transcription PCR 7.1 Protein Synthesis ■■ Multiplex PCR ■■ Bacterial Transcription ■■ Degenerate PCR ■■ Eukaryotic Transcription ■■ Nested PCR ■■ Translation ■■ Fast PCR ■■ Posttranslational Modifications ■■ Isothermal PCR 7.2 Protein Structure ■■ Random Amplification of Polymorphic DNA ■■ Amino Acids 6.5 PCR Optimization ■■ Protein Folding ■■ Quality of Template DNA 7.3 Proteins in Biology ■■ Primer Design ■■ Fibrous Proteins ■■ Cycling Parameters ■■ Globular Proteins ■■ Magnesium Concentration — Enzymes 7.4 Proteins in Biotechnology ■■ Proteins in Industry ■■ Proteins in Healthcare ■■ Proteins as Tools in Life Science Research

7.5 Methods of Protein Analysis Chapter 8: Immunological Applications ■■ Protein Quantitation 8.1 The Immune System — Biuret Test ■■ Cells of the Immune System — Lowry Assay ■■ Antibodies — Bradford Assay ■■ Immune Response ■■ Protein Properties Used in Analysis — Protein Size 8.2 Antibodies as Tools ■■ — Protein Charge Polyclonal Antibodies ■■ Monoclonal Antibodies ■■ Protein Mass Spectrometry ■■ Humanized Monoclonal Antibodies for Medical Use ■■ Protein Electrophoresis — Polyacrylamide Gels 8.3 Immunoassays — Electrophoresis Running Buffer ■■ Labeling and Detecting Antibodies — Discontinuous Buffer Systems ■■ Enzyme-Linked Immunosorbent Assay (ELISA) — Visualizing Proteins in Polyacrylamide Gels — ELISA for Direct Antigen Detection — ELISA for Indirect Antigen Detection ■■ Other Types of PAGE — Sandwich ELISA for Antigen Detection — Native PAGE — ELISA for Antibody Detection — Isoelectric Focusing — Instruments for ELISA — Two-Dimensional PAGE — Nucleic Acid Separation ■■ Ouchterlony Double Diffusion Assay ■■ Western Blotting ■■ Protein Chromatography — Electroblotting — Size Exclusion Chromatography — Immunodetection — Hydrophobic Interaction Chromatography — Western Blotting in Diagnostic Testing — Ion Exchange Chromatography — Affinity Chromatography ■■ Using Antibodies to Study Cells and Tissues — Analyzing the Results of Chromatography ■■ High-Throughput Immunoassays — Scaling Chromatography — Multiplex Bead Assays — Fluorescence-Activated Cell Sorting ■■ Protein Analysis Using Bioinformatics Chapter 8 Vignettes 7.6 Protein Production in Industry Bioethics — Testing for HIV Status at Home ■■ Protein Production ■■ Quality Control in Protein Production Biotech in the Real World — Using Immunodetection to Save the Panda Chapter 7 Vignettes Bioethics — Editing Genes to Cure Disease Biotech in the Real World — Car T Cell Therapy — A New Horizon in Cancer Treatment Biotech in the Real World — Vaccines in Bananas Careers in Biotech — Sophy Wong, MD, Associate Clinical Careers in Biotech — Scott Chilton, Marketing Manager, Professor of Medicine, University of California San Francisco, Maravai Life Sciences CA; Medical Director for HIV ACCESS and AETC How To... Set Up a Vertical Electrophoresis System How To... Select an Antibody Chapter 7 Activities Chapter 8 Activities 7.A Protein Quantitation Using the Bradford Assay 8.A Ouchterlony Double Immunodiffusion Assay 7.B Size Exclusion Chromatography 8.B Serum Antibody Detection by ELISA 7.C GFP Purification by Hydrophobic Interaction 8.C Quantitative ELISA Chromatography 8.D Western Blotting 7.D SDS-PAGE of Fish Muscle 7.E Biofuel Enzyme Assay 7.F Exploring Bioinformatics with GFP

Chapter 9: Research Projects 9.4 Project Ideas ■■ Research Tools and Techniques 9.1 What Is Research? ■■ Agricultural and Animal Research ■■ Deductive and Inductive Research ■■ Food Science ■■ Conducting Research in the Real World ■■ Humans and Health ■■ Peer Review ■■ Forensics ■■ Sharing of Scientific Information ■■ Energy and the Environment — Peer-Reviewed Publications — Conferences, Meetings, Seminars, and Posters 9.5 Considerations and Tips for All Laboratory Projects ■■ Microbiology 9.2 Student Research Projects ■■ Animal Research — Whole Class Projects ■■ Research on Human Subjects — Group Projects ■■ Molecular Biology — Individual Projects — Guidelines for Working with Recombinant DNA ■■ Collaborating with Scientists — Restriction Enzymes ■■ Venues for Conducting Research or — PCR Entering Competitions — Extracting DNA — High School Research — Primer Design — Undergraduate Research — Master Mixes

■■ Choosing a Research Project — Determining the Annealing Temperature ■■ Scoping a Research Project — Ligation and Transformation ■■ Planning a Project — DNA Sequencing ■■ Performing Background Research ■■ Proteins — Internet — Obtaining Antibodies — Libraries — Optimizing Immunoassays — Peer-Reviewed Journal Articles — Enzymes and Substrates — Mentors — Chromatography ■■ Experimental Design Chapter 9 Vignettes — Methods Bioethics — Clinical Trials and Data Fraud — Controls — Trials and Sample Size Biotech in the Real World — Regeneron Science Talent Search ■■ Data Analysis — Example of Data Analysis Careers in Biotech — Sunny Choe, PhD, Senior Medical — Calculating the Mean Scientist, Gilead Sciences, Inc. — Calculating the Standard Deviation How To... Read a Journal Article — Drawing Conclusions from Statistical Data Appendices ■■ Funding Research Projects A: Fast Gel Protocol 9.3 Communicating Research B: Alternative Staining Methods for Agarose Gels ■■ Posters ■■ Research Papers C: Stain-Free SDS-PAGE Gel Imaging ■■ Oral Presentations D: Glossary E: Laboratory Skills Assessment Rubric F: Laboratory Notebook and Project Rubrics

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