Introduction to experimental biophysics : a laboratory guide / Jay L. Nadeau. -- Boca Raton : CRC, Taylor & Francis Group, c2015. – (58.171057/N134)

Contents

Acknowledgment / vii

About the Author / ix

Preface: How to Use This Manual / xi

Introduction: Infrastructure and Safety Requirements / xv

Section One: Molecular Cloning

1 Transformation of Competent Bacteria with a DNA Plasmid / 3

2 Isolation of Plasmids by Miniprep and Restriction Enzyme Analysis / 15

3 Designing a New Construct Using Restriction Enzymes (Computer Lab) / 23

4 Restriction Enzymes / 33

5 Ligation / 41

6 Screening a Ligation and Choosing Recombinants / 53

8 Running the Polymerase Chain Reaction / 67

Section Two: Gene Expression

9 Protein Expression and Purification / 75

10 Characterization of Purified Protein / 83

11 Protein Crystallization / 89

Section Three: Fluorescent Reporter Genes in Mammalian Cells

12 Transfecting Mammalian Cells with Custom-Designed Constructs / 101

13 Screening Fluorescent Reporter Gene Expression in Mammalian Cells / 113

Section Four: Nanoparticles

14 Synthesis of Quantum Dots / 119

15 Synthesis of Gold Nanoparticles / 125

Section Five: Electrophysiology

16 Electrophysiology: Lipid Bilayer Recording / 131

17 Electrophysiology Alternative: Introduction to Lipid Vesicles / 137

Appendices: Instructor Information and Guides

Appendix 1 Disposables and Supplies Needed / 145

Appendix 2 Instructors' Guide to Behind-the-Scenes Preparation / 149

Appendix 3 Grading Recommendations and Guidelines / 153

Appendix 4 Advanced Options:

Customizing the Course / 163

Index / 165