Synthesis And Applications Of DNA And RNA

Synthesis And Applications Of DNA And RNA

1st Edition - May 7, 1987

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  • Editor: Saran Narang
  • eBook ISBN: 9780323158886

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Synthesis and Applications of DNA and RNA discusses the significant contributions in the development of synthetic routes to DNA and RNA. This book contains nine chapters that describe the complexities in the chemistry and biology of DNA and RNA. After briefly dealing with the various stages of development in the chemical synthesis of polynucleotides, this book goes on presenting the DNA synthesis on solid supports and through the phosphoramidite method on silica supports. The discussions then shift to the chemical-enzymatic synthesis of expressed genes; the biochemical aspects of chemical syntheses of oligoribonucleotides; and the methods of rapid DNA and RNA sequence analysis. A chapter specifically tackles the protocols of DNA synthesis using double-stranded plasmid DNA as a template. The final chapter deals with the use of oligonucleotides for the identification and isolation of specific gene sequences. This chapter also covers the use oligonucleotides in the detection of human genetic diseases. Biologists, geneticists, and researchers interested in DNA and RNA synthesis will find this work invaluable.

Table of Contents

  • Preface


    1. Development of Chemical Synthesis of Polynucleotides

    I. Introduction

    II. Historical Background

    III. Chemical Synthesis

    IV. Concluding Remarks


    2. DNA Synthesis on Solid Supports and Automation

    I. Introduction

    II. Mechanism of Coupling Reactions

    III. Polymer Supports

    IV. Protecting Groups Used during Oligonucleotide Synthesis

    V. Solid-Phase Oligonucleotide Synthesis

    VI. Postcoupling Reactions

    VII. Nonstandard Coupling Units

    VIII. Automation

    IX. Epilogue


    3. DNA Synthesis for Nonchemists: The Phosphoramidite Method on Silica Supports

    I. Introduction

    II. Solution-Phase Phosphite Triester Synthesis

    III. Solid-Phase Phosphite Triester Synthesis

    IV. Solid-Phase Phosphite Triester Synthesis of RNA

    V. Synthesis of DNA and RNA Analogs Using Phosphite Intermediates

    VI. Concluding Remarks


    4. Synthetic Gene Assembly, Cloning, and Expression

    I. Introduction

    II. General Methodology

    III. Reported Gene Syntheses

    IV. Conclusion


    5. Chemical Synthesis of RNA

    I. Introduction

    II. Protecting Groups and Condensing Reagents for Chemical Synthesis of Oligoribonucleotides

    III. Synthesis of Oligoribonucleotides by the Phosphotriester Method

    IV. Synthesis of Oligoribonucleotides by the Phosphite Method

    V. Enzymatic Joining of Oligoribonucleotides by RNA Ligase


    6. Rapid DNA Sequence Analysis

    I. Introduction

    II. Original Rapid DNA Sequencing Methods Developed between 1975 and 1977

    III. Improved Rapid Methods Developed between 1977 and 1982

    IV. Rapid Methods Involving Progressive Shortening of DNA

    V. Discussion


    7. Direct, Rapid RNA Sequence Analysis

    I. Introduction

    II. Preparation of the RNA Substrate

    III. Chemical Sequencing of RNA Molecules

    IV. Enzymatic Sequencing of RNA Molecules

    V. Probing RNA Conformation

    VI. Summary


    Appendix: Chemical Sequencing of End-Labeled RNA Molecules

    8. Oligonucleotide-Directed Site-Specific Mutagenesis Using Double-Stranded Plasmid DNA

    I. Introduction

    II. General Considerations

    III. Experimental Procedures

    IV. Remarks and Conclusions


    9. Synthetic Oligonucleotides for the Identification and Isolation of Specific Gene Sequences

    I. Introduction

    II. Early Studies

    III. Identification of Cloned DNA Sequences Based on the Primary Protein Sequence

    IV. Detection of Specific RNAs

    V. Detection of Specific Chromosomal Sequences

    VI. Detection of Human Genetic Diseases

    VII. Conclusion



Product details

  • No. of pages: 254
  • Language: English
  • Copyright: © Academic Press 1987
  • Published: May 7, 1987
  • Imprint: Academic Press
  • eBook ISBN: 9780323158886

About the Editor

Saran Narang

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