Methods in Virology

Methods in Virology

Volume V

1st Edition - December 28, 1971

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  • Editors: Karl Maramorosch, Hilary Koprowski
  • eBook ISBN: 9781483262239

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Methods in Virology, Volume V focuses on the methods used in virology, including hybridization, gel electrophoresis, freeze-etching technique, and ultracentrifugation. The selection first offers information on the fusion of cells for virus studies and production of cell hybrids; approaches to ultracentrifugation; and polyacrylamide gel electrophoresis of viral RNA. Discussions focus on applications to virological problems and analysis of results; analysis of the distribution of RNA on polyacrylamide gels after electrophoresis; and biological analysis of DNA components. The book then examines the polyacrylamide gel electrophoresis of viral proteins, DNA-RNA and DNA-DNA hybridization in virus research, and techniques of RNA-DNA hybridization in solution for the study of viral transcription. Topics include preparation of nucleic acids, hybridization and elution procedures that minimize RNA degradation, and procedures for acrylamide gel electrophoresis. The text takes a look at freeze-etching technique for the study of virus ultrastructure; procedures to increase virus yield from infected plants; and the immunoperoxidase technique. Concerns include principles of the immunoperoxidase technique, histochemical detection of peroxidase activity, sequence of events in virus infection, and factors affecting virus yield. The selection is a valuable source of data for researchers interested in the methods employed in virology.

Table of Contents

  • List of Contributors


    Contents of Other Volumes

    Chapter 1—Fusion of Cells for Virus Studies and Production of Cell Hybrids

    I. Introduction

    II. Methods

    III. Cell Fusion in Virology

    IV. Selection of Hybrid Cell Lines

    V. Uses of Heterokaryons and Hybrid Cell Lines


    Chapter 2—New Approaches to Ultracentrifugation

    I. Introduction

    II. A Solution: New Approaches

    III. Results

    IV. Discussion and Summary



    Chapter 3—Small Circular Viral DNA: Preparation and Analysis

    I. Introduction

    II. Virus Cultivation, Concentration, and Purification

    III. DNA Extraction and Purification

    IV. Physical Analysis of DNA Components

    V. Biological Analysis of DNA Components




    Chapter 4—Polyacrylamide Gel Electrophoresis of Viral RNA

    I. Introduction

    II. Apparatus

    III. Procedures for Preparation and Running of Gels

    IV. Analysis of the Distribution of RNA on Polyacrylamide Gels after Electrophoresis

    V. Elution of RNO from Analytical Gel Fractions

    VI. Large-Scale Preparative Techniques

    VII. Applications to Virological Problems and the Analysis of Results

    VIII. Conclusion



    Chapter 5—Polyacrylamide Gel Electrophoresis of Viral Proteins

    I. Introduction

    II. Procedures for Acrylamide Gel Electrophoresis

    III. Interpretation of Results

    IV. Discussion

    V. Appendix: Common Complaints, Troubleshooting Suggestions, and Tips


    Chapter 6—DNA-RNA and DNA-DNA Hybridization in Virus Research

    I. Introduction

    II. Materials

    III. DNA-RNA Hybridization Procedure

    IV. Characteristics of the DNA-RNA Hybridization Reaction

    V. Thermal Elution of RNA from RNA-DNA Hybrids

    VI. Hybridization and Elution Procedure that Minimizes RNA Degradation

    VII. DNA-DNO Hybridization

    VIII. Alternative Methods of Hybridization



    Chapter 7—Techniques of RNA-DNA Hybridization in Solution for the Study of Viral Transcription

    I. Introduction

    II. Basic Equipment

    III. Preparation of Nucleic Acids

    IV. RNA-DNA Hybridization in Solution (Liquid Hybridization)

    V. Segmental Transcription Studies by RNA-DNA Hybridization in Solution


    Chapter 8—Methods for the Study of mRNA Synthesis in Bacteriophage-Infected E. coli

    I. General Introduction

    II. Bacteriophage-Host System

    III. Isolation Methods

    IV. Analytical Methods



    Chapter 9—Reactions of Viruses in Agar Gel

    I. Introduction

    II. Reactions Between Viral Antigens and Antibiotics

    III. Reactions Between Viruses and Erythrocyte Stromata

    IV. Electron Microscopic Examination of Reaction Lines



    Chapter 10—Methods for the Study of Virus-Antibody Complexes

    I. General Introduction

    II. Methods of Reactivating Neutralized Virus

    III. Characterization of the Virus-Antibody Complex

    IV. Methods of Separating the Components of a Virus-Antibody Reaction Mixture


    Chapter 11—Techniques of Ferritin-Tagged Antibodies

    I. Introduction

    II. Purification of Ferritin

    III. Preparation of Antibody Globulin Fraction from Serum

    IV. Conjugation of Ferritin and Antibody

    V. Evaluating the Conjugate

    VI. Methods of Direct Ferritin-Tagging

    VII. Methods of Indirect Ferritin-Tagging

    VIII. Controls in Ferritin-Tagging Experiments

    IX. Hybrid Antibody Method for Ferritin Localization of Antigen

    X. Application of Hybrid Antibody

    XI. Other Methods of Visualizing Immune Globulins

    XII. Conclusions


    Chapter 12—The Immunoperoxidase Technique: Localization of Viral Antigens in Cells

    I. Introduction

    II. The Principles of the Immunoperoxidase Technique

    III. Properties of Horseradish Peroxidase

    IV. Histochemical Detection of Peroxidase Activity

    V. Peroxidase-Antibody Labeling Techniques

    VI. Procedure for Detection of Viral Antigens in Cells by the Immunoperoxidase Technique

    VII. Interpretation of the Results Obtained for the Localization of Viral Antigens in Cells by the Immunoperoxidase Technique



    Chapter 13—Leaf-Dip Serology

    I. Introduction

    II. Procedure

    III. Results and Discussion


    Chapter 14—Procedure to Increase Virus Yield from Infected Plants

    I. Introduction

    II. Sequence of Events in Virus Infection

    III. Criteria for Virus Yield

    IV. Factors Affecting Virus Yield

    V. Summary


    Chapter 15—Freeze-Etching Technique for the Study of Virus Ultrastructure

    I. Introduction

    II. The Freeze-Etching Technique

    III. Results up to the Present

    IV. Conclusion and Prospects



    Author Index

    Subject Index

Product details

  • No. of pages: 548
  • Language: English
  • Copyright: © Academic Press 1971
  • Published: December 28, 1971
  • Imprint: Academic Press
  • eBook ISBN: 9781483262239

About the Editors

Karl Maramorosch

Professor Karl Maramorosch works at Rutgers University, New Brunswick, NJ, USA.

Affiliations and Expertise

Rutgers University, New Brunswick, NJ, USA

Hilary Koprowski

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