Description
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 Cycle Douglas J. Dellinger, Jason R. Betley, Tadeusz K. Wyrzykiewicz, and Marvin H. Caruthers RNA Synthesis Using 2′-O-(Tert-Butyldimethylsilyl) Protection Brian S. Sproat RNA Oligonucleotide Synthesis Via 5′-Silyl-2′-Orthoester Chemistry Stephanie A. Hartsel, David E. Kitchen, Stephen A. Scaringe, and William S. Marshall Dimethylthiarum Disulfide: New Sulfur Transfer Reagent in Phosphorothioates Oligonucleotide Synthesis Zhiwei Wang, Quanlai Song, and Yogesh S. Sanghvi Assay for Evaluating Ribonuclease H-Mediated Degradation of RNA-Antisense Oligonucleotide Duplexes Annie Galarneau, Kyung-Lyum Min, Maria M. Mangos, and Masad J. Damha Di- and Oligonucleotide Synthesis Using H-Phosphonate Chemistry Jacek Stawinski and Roger Strmberg Solid-Phase Synthesis of Circular Oligonucleotides Enrique Pedroso, Nuria Escaja, Miriam Frieden, and Anna Grandas Locked Nucleic Acid Synthesis Henrik M. Pfundheller, Anders M. Srensen, Christian Lomholt, Anders M. Johansen, Troels Koch, and Jesper Wengel Synthesis of DNA Mimics Representing HypNA-pPNA Hetero-oligomers Vladimir A. Efimov and Oksana G. Chakhmakhcheva Base-Modified Oligonucleotides With Increased Duplex Stability: Pyrazolo[3,4-d]pyrimidines Replacing Purines Frank Seela, Yang He, Junlin He, Georg Becher, Rita Krschel, Matthias Zulauf, and Peter Leonard Introduction of Hypermodified Nucleotides in RNA Darrell R. Davis and Ashok C. Bajji Chemical Methods for Peptide-Oligonucleotide Conjugate Synthesis Dmitry A. Stetsenko and Michael J. Gait Biotin-Labeled Oligonucleotides With Extraordinarily Long Tethering Arms A. Michael Morocho, Valeri Karamyshev, Olga Shcherbinina, and Nikolai Polushin Universal Labeling Chemistry for Nucleic Acid Detection on DNA Chips Ali Laayoun, Eloy Bernal-Mndez, Isabelle Sothier, Mitsuharu Kotera, and Alain Troesch Postsynthetic Functionalization of Triple Helix-Forming Oligonucleotides Alexandre S. Boutorine, and Jian-Sheng Sun Fluorescence-Based On-Line Detection as an Analytical Tool in RNA Electrophoresis Ute Scheffer and Michael Gbel Design and Optimization of Molecular Beacon Real-Time Polymerase Chain Reaction Assays Jacqueline A. M. Vet and Salvatore A. E. Marras High-Throughput Production of Optimized Primers (Fimers) for Whole-Genome Direct Sequencing Nikolai Polushin, Andrei Malykh, A. Michael Morocho, Alexei Slesarev, and Sergei Kozyavkin Nonenzymatic Template-Directed RNA Synthesis Marcus Hey and Michael Gbel DNAse I Footprinting of Small Molecule Binding Sites on DNA Christian Bailly, Jrme Kluza, Christopher Martin, Thomas Ellis, and Michael J. Waring Gene Targeting Using Peptide Nucleic Acid Peter E. Nielsen Nucleic Acid Library Construction Using Synthetic DNA Constructs Hani S. Zaher and Peter J. Unrau In Vitro Selection From Combinatorial Nucleic Acid Libraries Andres Jschke In Vitro Selection Procedures for Identifying DNA and RNA Aptamers Targeted to Nucleic Acids and Proteins Eric Dausse, Christian Cazenave, Bernard Rayner, and Jean-Jacques Toulm Short Double-Stranded Ribonucleic Acid as Inhibitor of Gene Expression by the Interference Mechanism Jean-Rmi Bertrand, Andrei Maksimenko, and Claude Malvy Index
