Basic Techniques for the Biochemical Laboratory

Chapter 1 Introduction to the Biochemistry Laboratory 
1.1 Laboratory Safety 
1.2 The Importance of Keeping a Laboratory Notebook 
1.3 Working in Groups 
1.4 Writing Appropriate Lab Reports 
1.5 Micropipetting Techniques and Calibration 
1.6 A Refresher on Dilution of Concentrated Solutions 
1.7 Experimental Details 

Chapter 2 Spectroscopic Techniques in the Biochemistry Laboratory 
2.1 A Refresher on pH of Aqueous Solutions and Buffers 
2.2 Using UV-Visible Spectroscopy to Determine the pKa of a pH Indicator Dye 
2.3 The Origin of the Beer–Lambert Law 
2.4 Experimental Details 

Chapter 3 Protein Quantitation Using the Bradford Assay 
3.1 The Chemistry of the Bradford Assay 
3.2 UV-Visible Spectral Changes upon Protein Binding by the Bradford Reagent 
3.3 Creating a Standard Curve with Bovine Serum Albumin 
3.4 Linear Curve Fitting and Determination of Unknown Protein Concentrations
3.5 Experimental Details 

Chapter 4 Size Exclusion Chromatography (SEC) for Protein Purification
4.1 Introduction to Size Exclusion Chromatography
4.2 Structure of SEC Media
4.3 Basics of Pouring SEC Columns
4.4 Separation of a Mixture By Size
4.5 A Simple Method for Determining the Relative Mobility of Chromatic Molecules on an SEC Column 
4.6 Experimental Details

Chapter 5 Protein Electrophoresis
5.1 Fundamentals of Object Mobility in Electric Fields
5.2 Chemical Properties of Polyacrylamide Gels for Protein Electrophoresis 
5.3 Assembly of Electrophoresis Chambers 
5.4 Creating a Standard Curve from Relative Mobilities of Proteins and Determination of Molecular Weights of Unknown Proteins
5.5 Experimental Details 

Chapter 6 Plasmid DNA Transformation of Bacteria
6.1 The Basics of Plasmid DNA 
6.2 Heat-Shock Transformation of Bacteria
6.3 Storage of Transformed bacteria
6.4 Experimental Details 

Chapter 7 Green Fluorescent Protein Purification by Hydrophobic Interaction Chromatography (HIC)
7.1 Introduction to Hydrophobic Interaction Chromatography (HIC)
7.2 Properties of Green Fluorescent Protein
7.3 Preparation of E. Coli Lysate for GFP Purification
7.4 Using UV Light to Visualize Movement of GFP in HIC Media 
7.5 Experimental Details 

Chapter 8 DNA Electrophoresis and Restriction Nucleases
8.1 Electrophoretic Mobility Revisited
8.2 Properties of Agarose Gels
8.3 Using Restriction Nucleases to Cleave DNA
8.4 Determination of DNA Fragment Sizes
8.5 Experimental Details 

Chapter 9 Ligand Binding to Biomacromolecules
9.1 A Refresher on the Principles of Chemical Equilibrium and Gibbs Free Energy
9.2 Derivation of the Scatchard Plot for Ligand–Macromolecule Binding 
9.3 Alternatives to the Scatchard Plot: Nonlinear Curve Fitting
9.4 The Use of Spectroscopic Techniques to Measure Binding Affinities
9.5 Experimental Details 

Chapter 10 Enzyme Kinetics
10.1 A Refresher on the Principles of Catalysis
10.2 The Michaelis–Menten Model of Enzyme Activity
10.3 The Lineweaver–Burk Plot
10.4 The Use of Spectroscopic Techniques to Measure Initial Velocities
10.5 Experimental Details

Chapter 11 The Polymerase Chain Reaction (PCR)
11.1 The Basics of PCR 
11.2 Programming the Thermocycler
11.3 The Use of PCR in Criminal Investigations
11.4 Experimental Details 

Chapter 12 Bioinformatics and Molecular Graphics Techniques
12.1 The Dawn of Molecular Biology 
12.2 The Research Collaboratory for Structural Bioinformatics Protein Data Bank (RCSB PDB)
12.3 Finding Structures in the RCSB PDB
12.4 Analyzing Structures in the RCSB PDB
12.5 Experimental Details: Understanding Enzyme-Active Sites and Dna Structure Using the RCSB PDB