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Methods in Virology - 1st Edition - ISBN: 9780124702059, 9781483262239

Methods in Virology

1st Edition

Volume V

Editors: Karl Maramorosch Hilary Koprowski
eBook ISBN: 9781483262239
Imprint: Academic Press
Published Date: 28th December 1971
Page Count: 548
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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


No. of pages:
© Academic Press 1971
28th December 1971
Academic Press
eBook ISBN:

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

Ratings and Reviews