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Fundamental Laboratory Approaches for Biochemistry and Biotechnology 2e / Edition 2

Fundamental Laboratory Approaches for Biochemistry and Biotechnology 2e / Edition 2


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Fundamental Laboratory Approaches for Biochemistry and Biotechnology 2e / Edition 2

Ninfa/Ballou/Benore is a solid biochemistry lab manual, dedicatedto developing research skills in students, allowing them to learntechniques and develop the organizational approaches necessary toconduct laboratory research.

Ninfa/Ballou/Benore focuses on basic biochemistry laboratorytechniques with a few molecular biology exercises, a reflection ofmost courses which concentrate on traditional biochemistryexperiments and techniques. The manual also includes anintroduction to ethics in the laboratory, uncommon in similarmanuals. Most importantly, perhaps, is the authors' three-prongedapproach to encouraging students to think like a researchscientist: first, the authors introduce the scientific method andthe hypothesis as a framework for developing conclusiveexperiments; second, the manual's experiments are designed tobecome increasingly complex in order to teach more advancedtechniques and analysis; finally, gradually, the students arerequired to devise their own protocols. In this way, students andinstructors are able to break away from a "cookbook" approach andto think and investigate for themselves.

Suitable for lower-level and upper-level courses; Ninfa spansthese courses and can also be used for some first-year graduatework.

Product Details

ISBN-13: 9780470087664
Publisher: Wiley
Publication date: 03/23/2009
Pages: 480
Sales rank: 312,500
Product dimensions: 8.40(w) x 10.80(h) x 0.70(d)

Table of Contents

Chapter 1: Getting Started in Scientific Research.

1.1 The Difference Between Experiments and Demonstration.

1.2 Philosophy and Design of Experiments.

1.3 Designing Informative Experiments.

1.4 Ethics in Science.

1.5 Keeping a Laboratory Notebook.

1.6 Laboratory Reports.

1.7 Presentation and Analysis of Data.

1.8 The Minisymposium.

Chapter 2: Basic Procedures in the BiochemistryLaboratory.

2.1 Laboratory Safety.

2.2 Special Safety Procedures Are Required for Using RadioactiveMaterials and Operating the Autoclave.

2.3 Measurement of Weights, Volumes, and pH.

2.4 Various Instruments Used.

2.5 Other General Techniques.

2.6 Solutions and Dilutions.

2.7 Buffers and pH.

2.8 Appendix Calculating Titration Curves for Polyprotic Acidsand Other Multiple Binding Site Receptors.

2.9 Equipment Used in This Course.

Chapter 3: Spectroscopic Methods.

3.1 Introduction.

3.2 Design and Properties of Spectrophotometers.

3.3 Effects of Spectral Bandpass and Stray Light.

3.4 Recording Spectrophotometers.

3.5 Fluorescence Spectroscopy.

3.6 Chromogenic and Fluorogenic Reactions Used for Analysis.

3.7 Other Spectroscopic Techniques.

3.8 Mass Spectrometry (MS).

Experiments 3-1 to 3-3.

Reagents Needed for Chapter 3.

Chapter 4: Quantification of Protein Concentration.

4.1 Purposes of Protein Quantification.

4.2 Factors to Consider in Choosing an Assay.

4.3 Non-Colorimetric Procedures for Quantification ofProteins.

4.4 Colorimetric Procedures for Quantification of Proteins.

Experiment 4-1.

Reagents Needed for Chapter 4.

Chapter 5: Chromatography.

5.1 Introduction.

5.2 Gel-Filtration (Size Exclusion or Gel-Permeation)Chromatography.

5.3 Affinity Chromatography.

5.4 Ion-Exchange Chromatography.

5.5 Hydrophobic Interaction Chromatography.

Experiments 5-1 and 5-2.

Reagents Needed for Chapter 5.

Chapter 6: Gel Electrophoresis of Proteins.

6.1 Process of Electrophoresis.

6.2 Polyacrylamide Gels.

6.3 SDS-Polyacrylamide Gel Electrophoresis (SDS-PAGE) ofProteins.

6.4 Detection of Proteins in SDS-Polyacrylamide Gels.

6.5 Applications of SDS-PAGE.

Experiments 6-1 and 6-2.

Reagents Needed for Chapter 6.

Chapter 7: Overview of Protein Purification.

7.1 Introduction.

7.2 Development of a Suitable Assay Procedure.

7.3 Time, Temperature, and Yield.

7.4 Selection of the Best Source Material.

7.5 Solubilization of the Protein.

7.6 Initial Steps of Purification.

7.7 Developing a Series of High-Resolution ChromatographicSteps.

7.8 Methods Used to Change Buffer and Concentrate ProteinSamples.

7.9 A Logical Series of Steps.

7.10 Storage of the Purified Protein.

7.11 The Protein Purification Table.

Chapter 8: Subcellular Fractionation.

8.1 Introduction.

8.2 Structural Organization of Prokaryotic and EukaryoticCells.

8.3 Overview of Fractionation Protocols.

Reagents and Equipment Needed for Chapter 8.

Chapter 9: Isolation and Characterization of TheEnzyme-alkaline phosphatase from EscHerichia Coli.

9.1 Objectives.

9.2 Introduction and Basic Principles.

9.3 Purification of Alkaline Phosphatase.

9.4 Characterization of Purified Alkaline Phosphatase.

Appendix 9-1 Assay of Alkaline Phosphatase.

Reagents and Equipment Needed for Chapter 9.

Chapter 10: Enzyme Kinetics.

10.1 Why Use Steady-State Kinetics?

10.2 Steady-State Kinetics Principles.

10.3 The Significance of Km and Vmax.

10.4 Graphical Analysis.

10.5 Competitive, "Noncompetitive," and "Uncompetitive"Inhibitors.

Experiments 10-1 to 10-3.

Reagents Needed for Chapter 10.

Chapter 11: Ligand Binding.

11.1 Ligand Binding Is the Key to Most Biological Processes.

11.2 Analysis of Ligand Binding at Equilibrium.

11.3 Digression on Regression.

11.4 Effects of the Concentrations of L and R.

11.5 Effects of Two Sites and Cooperative Behavior.

11.6 Analysis of the Kinetics of Ligand Binding.

11.7 Methods Used to Study Receptor-Ligand Interactions.

Experiments 11-1 and 11-2.

Reagents Needed for Chapter 11.

Chapter 12: Enzymatic Methods of Analysis.

12.1 Enzymatic Analysis of Substrates.

12.2 Assays for Enzymatic Activity.

12.3 Practical Considerations.

12.4 Coupled Assays.

12.5 Experiments with Pyridine Nucleotide-Requiring Enzymes.

Experiments 12-1 to 12-4.

Reagents Needed for Chapter 12.

Appendix 12-1: Sample Calculations.

Chapter 13: Recombinant DNA Techniques.

13.1 Introduction.

13.2 Properties of Nucleic Acids.

 13.3 Strategy of Recombinant DNA Techniques.

13.4 Cutting and Splicing DNA.

13.5 Gel Electrophoresis of DNA.

13.6 Introducing DNA into Cells.

13.7 Identifying Transformed Cells.

13.8 Vectors, Hosts, and Libraries for Recombinant DNAExperiments.

13.9 Applications of Recombinant DNA Technology.

Experiments 13-1 to 13-4.

Reagents Needed for Chapter 13.

Chapter 14: Polymerase Chain Reaction (PCR)Technology.

14.1 Introduction.

14.2 Principle of the PCR Method.

14.3 The Three Steps of PCR Are Controlled by Temperature.

14.4 Applications of PCR.

14.5 Legal and Ethical Issues.

Experiments 14-1 to 14-6.

Reagents Needed for Chapter 14.

Chapter 15: Using the Computer and the Internet forBiochemical Research and Communication.

15.1 Introduction to the World Wide Web (the Internet).

15.2 Information on the Internet Useful to Biochemists.

15.3 Literature and Data Searches.

15.4 Visualization of Molecular Structures.

15.5 Other Useful Information.

15.6 Homework Assignment.

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