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Synthesis And Applications Of DNA And RNA

1st Edition - May 7, 1987

Editor: Saran Narang

eBook ISBN:

9 7 8 - 0 - 3 2 3 - 1 5 8 8 8 - 6

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… Read more

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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.

Preface

Introduction

1. Development of Chemical Synthesis of Polynucleotides

I. Introduction

II. Historical Background

III. Chemical Synthesis

IV. Concluding Remarks

References

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

References

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

References

4. Synthetic Gene Assembly, Cloning, and Expression

I. Introduction

II. General Methodology

III. Reported Gene Syntheses

IV. Conclusion

References

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

References

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

References

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

References

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

References

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

References

Index