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 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