Oligonucleotide Synthesis Methods and Applications 1st Edition by Piet Herdewijn 9781588292339 1588292339

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Product details:

ISBN 10: 1588292339

ISBN 13: 9781588292339

Author: Piet Herdewijn

A collection of powerful new techniques for oligonucleotide synthesis and for the use of modified oligonucleotides in biotechnology. Among the protocol highlights are a novel two-step process that yields a high purity, less costly, DNA, the synthesis of phosphorothioates using new sulfur transfer agents, the synthesis of LNA, peptide conjugation methods to improve cellular delivery and cell-specific targeting, and triple helix formation. The applications include using molecular beacons to monitor the PCR amplification process, nuclease footprinting to study the sequence-selective binding of small molecules of DNA, nucleic acid libraries, and the use of small interference RNA (siRNA) as an inhibitor of gene expression.

Table of contents:

1. Synthesis of DNA Using a New Two-Step Cycle

2. RNA Synthesis Using 2′-O-(Tert-Butyldimethylsilyl) Protection

3. RNA Oligonucleotide Synthesis Via 5′-Silyl-2′-Orthoester Chemistry

4. Dimethylthiarum Disulfide

5. Assay for Evaluating Ribonuclease H-Mediated Degradation of RNA-Antisense Oligonucleotide Duplexes

6. Di- and Oligonucleotide Synthesis Using H-Phosphonate Chemistry

7. Solid-Phase Synthesis of Circular Oligonucleotides

8. Locked Nucleic Acid Synthesis

9. Synthesis of DNA Mimics Representing HypNA-pPNA Hetero-Oligomers

10. Base-Modified Oligonucleotides With Increased Duplex Stability

11. Introduction of Hypermodified Nucleotides in RNA

12. Chemical Methods for Peptide-Oligonucleotide Conjugate Synthesis

13. Biotin-Labeled Oligonucleotides With Extraordinarily Long Tethering Arms

14. Universal Labeling Chemistry for Nucleic Acid Detection on DNA Chips

15. Postsynthetic Functionalization of Triple Helix-Forming Oligonucleotides

16. Fluorescence-Based On-Line Detection as an Analytical Tool in RNA Electrophoresis

17. Design and Optimization of Molecular Beacon Real-Time Polymerase Chain Reaction Assays

18. High-Throughput Production of Optimized Primers (Fimers) for Whole-Genome Direct Sequencing

19. Nonenzymatic Template-Directed RNA Synthesis

20. DNase I Footprinting of Small Molecule Binding Sites on DNA

21. Gene Targeting Using Peptide Nucleic Acid

22. Nucleic Acid Library Construction Using Synthetic DNA Constructs

23. In Vitro Selection From Combinatorial Nucleic Acid Libraries

24. In Vitro Selection Procedures for Identifying DNA and RNA Aptamers Targeted to Nucleic Acids and Proteins

25. Short Double-Stranded Ribonucleic Acid as Inhibitor of Gene Expression by the Interference Mechanism

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Tags: Piet Herdewijn, Oligonucleotide, Synthesis, Methods