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.
Sprache
Verlagsort
Verlagsgruppe
Elsevier Science Publishing Co Inc
Zielgruppe
Für höhere Schule und Studium
Für Beruf und Forschung
Gewicht
ISBN-13
978-0-12-514030-0 (9780125140300)
Copyright in bibliographic data is held by Nielsen Book Services Limited or its licensors: all rights reserved.
Schweitzer Klassifikation
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
