Synthesis And Applications Of DNA And RNA

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.

Contenu

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