Basic Bioscience Laboratory Techniques E-Book

Philip L. R. Bonner

• Herausgeber: John Wiley & Sons
• Kategorie: Wissenschaft und neue Technologien
• Sprache: Englisch

Seitenzahl: 284

Veröffentlichungsjahr: 2011

Beschreibung

This unique, practical, pocket-sized guide and reference provides every first year bioscience student with all they need to know to prepare reagents correctly and perform fundamental laboratory techniques. It also helps them to analyse their data and present their findings, in addition to directing the reader, via a comprehensive list of references, to relevant further reading All of the core bioscience laboratory techniques are covered including: basic calculations and the preparation of solutions; aseptic techniques; microscopy techniques; cell fractionation ; spectrophotometry; chromatography of small and large molecules: electrophoresis of proteins and nucleic acids and data analysis. In addition the book includes clear, relevant diagrams and worked examples of calculations. In short, this is a 'must-have' for all first year bioscience students struggling to get to grips with this vitally important element of their course.

Leseprobe

Contents

Preface

Glossary

Abbreviations

1 THE PREPARATION OF SOLUTIONS IN BIOSCIENCE RESEARCH

1.1 Introduction

1.2 Concentration

1.3 Using Balances to Weigh Out Reagents

1.4 Practical Considerations When Making a 1.0 M Solution

1.5 Dilutions and the Use of Pipettes

1.6 Water, Acids and Bases

1.7 Buffers

1.8 The Equilibrium/Dissociation Constant (Ka) for an Acid or Base and the Henderson –Hasselbalch Equation

1.9 Summary

Notes

2 MICROSCOPY

2.1 Introduction

2.2 Microscopes – General Principles

2.3 Principles of Image Formation

2.4 Light Microscopy

2.5 Electron Microscopy

2.6 Summary

3 SPECTROPHOTOMETRY

3.1 Introduction

3.2 The Electromagnetic Spectrum

3.3 The Absorbance of Light

3.4 Absorption Spectrophotometry

3.5 The Laws of the Absorbance of Light

3.6 The Beer Lambert Law

3.7 Spectrophotometers

3.8 Applications of Spectrophotometry in Bioscience

3.9 Summary

Notes

4 DATA ANALYSIS AND PRESENTATION

4.1 Introduction

4.2 Statistical Analysis of Data: Some Key Definitions

4.3 Distributions

4.4 Statistical Comparison of Data

4.5 Presentation, Structure and Organization of Data in Laboratory Reports

4.6 Summary

5 THE EXTRACTION AND CLARIFICATION OF BIOLOGICAL MATERIAL

5.1 General Introduction

5.2 Extraction

5.3 Extraction Methods for Animal and Plant Tissue

5.4 Extraction Methods for Bacteria

5.5 Clarification

5.6 Centrifugation Techniques

5.7 Points of Good Practice in Centrifugation

5.8 Summary

Notes

6 ELECTROPHORESIS OF PROTEINS AND NUCLEIC ACIDS

6.1 General Introduction

6.2 Separation of Protein Mixtures by Gel Electrophoresis

6.3 Other Electrophoretic Techniques Applied to Proteins

6.4 Separation of Nucleic Acids by Gel Electrophoresis

6.5 Applications of Gel Electrophoresis of Nucleic Acids

6.6 Summary

7 CHROMATOGRAPY

7.1 General Introduction

7.2 The Theory of Chromatography

7.3 Factors to Consider in Chromatography

7.4 Methods Used to Elute Samples in Chromatography

7.5 Different Types of Chromatography and What Properties Can be Used to Separate Molecules

7.6 Thin Layer Chromatography (TLC)

7.7 High Pressure Liquid Chromatography (HPLC)

7.8 Gas Liquid Chromatography (GLC)

7.9 Ion Exchange Chromatography (IEX)

7.10 Size Exclusion Chromatography

7.11 Affinity Chromatography

7.12 Summary

Notes

8 CELL CULTURE TECHNIQUES

8.1 Introduction

8.2 Growth and Maintenance of Cells in Culture

8.3 Summary

Notes

Suggestions for further reading

Index

This edition first published 2011 © 2011 by John Wiley & Sons, Ltd.

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Library of Congress Cataloguing-in-Publication Data

Bonner, Philip L. R.

Basic bioscience laboratory techniques: a pocket guide/Philip Bonner and Alan Hargreaves.

p.; cm.

Includes bibliographical references and index.

ISBN 978-0-470-74309-6 (pbk.)

1. Biology—Laboratory manuals. 2. Biology–Handbooks, manuals, etc. I. Hargreaves, Alan. II. Title.

[DNLM: 1. Laboratory Techniques and Procedures–Handbooks. 2. Biotechnology–Handbooks. 3. Research-Handbooks. QY 39]

QH317.B66 2010

570.78–dc22

2010033321

PREFACE

In recent years there have been many advances made in the field of bioscience research. This progress has been driven by parallel advances in methodology. New and minor modifications to existing techniques can propel and enhance scientific investigation. However, the use of any procedure can only be advantageous if there is an appreciation of the basic principles, requirements, benefits and pitfalls of the techniques employed. This is true for both experienced as well as novice investigators.

We have been teaching in the HE sector for the past 20 years. During this period we have become increasingly aware that newly enrolled students are struggling to appreciate the basics of laboratory techniques and data analysis. Although the reasons for this are many fold, a sound platform for scientific investigation is still dependent upon the understanding and correct use of basic bioscience laboratory techniques. Our students perceive that there is a lack of appropriate support in the literature. With this in mind, the rationale for this book is to supply you (students) with an easy to use reference book that will provide you with a basis for understanding the basic bioscience techniques that you will come across in your HE studies.

The book provides the foundation of many basic laboratory techniques, worked examples of common calculations and (we hope) helpful tips. We wanted the book to be used prior to, during and after laboratory classes, which is why it is designed to fit into the pocket of a laboratory coat. The content reflects the size constraints and accompanying material will be available on a companion web page. In addition, supplementary reading will be needed to provide more detailed information on the techniques covered. However, we believe that the book will be of benefit to students starting their degree courses in any biosciences theme. The techniques covered in this book will be encountered by you repeatedly during laboratory sessions and we hope that the book will remain a valuable reference text throughout your degree course.

We would both like to thank our families (Alan: Olivia, Alan, Elena and Stephanie. Phil: Elizabeth, Francesca, Mark and Neil) and friends (Chris, David, John, Krishan, Lee, Mel and Wayne) for their support and suggestions during the preparation of this book. We would also like to thank Nicky and Fiona at Wiley and Sons for their patience and encouragement during the book’s preparation period.

Philip L.R. BonnerAlan J. Hargreaves

ABBREVIATIONS

A260nm: Absorbance at 260 nm AA34o nm: The change in absorbance at 340 nm A: Adenine ATCC: American Type Culture Collection ATP: Adenosine 5’-triphosphate AMPS: Ammonium persulphate Bp: base pair C: Cytosine C18: a chain of 18 carbon atoms C8: a chain of 8 carbon atoms C5: a chain of 5 carbon atoms CHAPS: (3-[(3-Cholamidopropyl) dimethylammonio]-1-propane sulphonate