Recombinant DNA Laboratory Manual

Judith W. Zyskind, Sanford I. Bernstein

• Publisher: Academic Press
• Year: 2014
• ISBN:
  9781483220970
• Topic: Biological Sciences
• Subtopic: Molecular Biology
• Index: Biological Sciences

Recombinant DNA Laboratory Manual is a laboratory manual on the fundamentals of recombinant DNA techniques such as gel electrophoresis, in vivo mutagenesis, restriction mapping, and DNA sequencing. Procedures that are useful for studying either prokaryotes or eukaryotes are discussed, and experiments are included to teach the fundamentals of recombinant DNA technology. Hands-on computer sessions are also included to teach students how to enter and manipulate sequence information. Comprised of nine chapters, this book begins with an introduction to bacterial growth parameters, how to measure bacterial cell growth, and how to plot cell growth data. The discussion then turns to the isolation and analysis of chromosomal DNA in bacteria and Drosophila; plasmid DNA isolation and agarose gel analysis; and introduction of DNA into cells. Subsequent chapters deal with Tn5 mutagenesis of pBR329; DNA cloning in M13; DNA sequencing; and DNA gel blotting, probe preparation, hybridization, and hybrid detection. The book concludes with an analysis of lambda phage manipulations. This manual is intended for advanced undergraduate or beginning graduate students and should also be helpful to established investigators who are changing their research focus.

Table of contents

• Front Cover
• Recombinant DNA Laboratory Manual
• Copyright Page
• Table of Contents
• Dedication
• PREFACE
• SCHEDULE OF LABORATORY EXERCISES
• CHAPTER 1. LAB I. BACTERIAL GROWTH PARAMETERS
• CHAPTER 2. LAB II. ISOLATION AND ANALYSIS OF BACTERIAL AND DROSOPHILA CHROMOSOMAL DNA
• CHAPTER 3. LAB III. PLASMID DNA ISOLATION AND AGAROSE GEL ANALYSIS
• CHAPTER 4. Lab IV. INTRODUCTION OF DNA INTO CELLS
• CHAPTER 5. LAB V. TN5 MUTAGENESIS OF PBR329
• CHAPTER 6. LAB VI. DNA CLONING IN M13
• CHAPTER 7. LAB VII. DNA SEQUENCING
• CHAPTER 8. LAB VIII DNA GEL BLOTTING, PROBE PREPARATION, HYBRIDIZATION, AND HYBRID DETECTION
• CHAPTER 9. LAB IX LAMBDA PHAGE MANIPULATIONS
• Appendix 1: USE OF COMPUTERS IN A MOLECULAR BIOLOGY LABORATORY
• Appendix 2: SAFETY IN THE RECOMBINANT DNA LABORATORY
• Appendix 3: SOLUTIONS
• Appendix 4: STRAINS AND DNA
• Appendix 5: MOLECULAR BIOLOGY REAGENT SUPPLIERS
• Appendix 6: FREQUENTLY USED ENZYMES
• Appendix 7: FREQUENTLY USED TECHNIQUES NOT DESCRIBED IN LABS
• INDEX