Description

Basic Methods in Molecular Biology discusses the heart of the most recent revolution in biology—the development of the technology of genetics. The achievements in this field have simply changed what biologists do and, perhaps even more important, the way they think. Moreover, never before have scientists from such a broad range of disciplines rushed into such a small and slightly arcane field to learn and carry off a bit of the technology. This book comprises 21 chapters, opening with three introductory ones that discuss the basics of molecular biology; the tools of the molecular biologist; and general preparations, procedures, and considerations for use of the book. The following chapters then discuss cloning vectors and bacterial cells; preparation of DNA from eukaryotic cells; probing nucleic acids; plasmid DNA preparation; DNA restriction fragment preparation; purification of DNA; and preparation and analysis of RNA from eukaryotic cells. Other chapters cover preparation of DNA from bacteriophage clones; cloning DNA from the eukaryotic genome; subcloning into plasmids; M13 cloning and sequencing; further characterization of cloned DNA; transfection of mammalian cells in culture; protein methods; general methods; and specialized methods. This book will be of interest to practitioners in the fields of biology and molecular genetics.

Table of Contents

Foreword Acknowledgments 1. The Basics of Molecular Biology 2. The Tools of the Molecular Biologist 3. General Preparations, Procedures, and Considerations for Use of Manual Section 3-1 Using This Manual 3-2 Safety Considerations 3-3 Equipment Needed for Molecular Biology Studies 4. Cloning Vectors and Bacterial Cells Section 4-1 pBR322 4-2 M13 4-3 pUC 4-4 λgtlO 4-5 λgtll 4-6 EMBL3 and EMBL4 4-7 Charon 28 4-8 Bacterial Strains 5. Preparation of DNA from Eukaryotic Cells Section 5-1 Rapid DNA Preparation 5-2 Preparation of DNA from Eukaryotic Cells: General Method 5-3 DNA Preparation from Cultured Cells and Tissue 5-4 Restriction Endonucleases (REs) and Their Use 5-5 Agarose Gel Electrophoresis 5-6 Southern Blot 6. Probing Nucleic Acids with Labeled Synthetic Probes Section 6-1 Making Synthetic DNA Probes: General Description 6-2 End Labeling of Synthetic Probes 6-3 Hybridization with Synthetic 32P End-Labeled Probe 7. Probing Nucleic Acids with Plasmid-Derived Probes Section 7-1 Nick Translation 7-2 DNA Hybridization (Southern Blot Hybridization) 8. Plasmid DNA Preparation Section 8-1 Transformation of Bacteria 8-2 Plasmid DNA Preparation: Triton-Lysozyme Method 8-3 Large-Scale Alkaline Lysis Method: Plasmid Purification 8-4 Plasmid "Mini-Prep" Method 9. DNA Restriction Fragment Preparation Section 9-1 Minigels 1 9-2 Analysis of DNA Fragments After Enzymatic Cleavage: Agarose Gel Electrophoresis 9-3 Electroelution 9-4 Polyacrylamide Gel Electrophoresis of DNA Restriction Fragments 10. Purification of DNA Section 10-1 Spermine Purification of DNA

Details

No. of pages:
399
Language:
English
Copyright:
© 1986
Published:
Imprint:
Elsevier
Print ISBN:
9780444010827
Electronic ISBN:
9780444601490