Oligonucleotide Synthesis: Methods and Applications [Paperback]

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.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.Synthesis of DNA Using a New Two-Step CycleDouglas J. Dellinger, Jason R. Betley, Tadeusz K. Wyrzykiewicz, and Marvin H. CaruthersRNA Synthesis Using 2'-O-(Tert-Butyldimethylsilyl) ProtectionBrian S. SproatRNA Oligonucleotide Synthesis Via 5'-Silyl-2'-Orthoester ChemistryStephanie A. Hartsel, David E. Kitchen, Stephen A. Scaringe, and William S. MarshallDimethylthiarum Disulfide: New Sulfur Transfer Reagent in Phosphorothioates Oligonucleotide SynthesisZhiwei Wang, Quanlai Song, and Yogesh S. SanghviAssay for Evaluating Ribonuclease H-Medlăg