Methods in Molecular Biology 288Oligonucleotide Synthesis [1ed.]9781588292339, 1588292339, 1592598234
Description:Nucleic acids chemistry has progressed rapidly in the last 10 years with the introduction of many new techniques and the development of many novel therapeutic entities. In Oligonucleotide Synthesis: Methods and Applications, laboratory experts describe in step-by-step detail the powerful new techniques they have developed for oligonucleotide synthesis and for the use of modified oligonucleotides in biotechnology. Among the protocol highlights are the synthesis of phosphorothioates via new sulfur transfer agents, the synthesis of LNA, peptide conjugation methods to improve cellular delivery and cell-specific targeting, triple helix formation, and a novel two-step process that yields a high purity DNA that is less costly. Novel applications include the use of molecular beacons to monitor the PCR amplification process, nuclease footprinting to study the sequence-selective binding of small molecules to DNA, nucleic acid libraries, optimization of RNA synthesis for siRNA applications, and the use of small interference RNA (siRNA) as an inhibitor of gene expression. The protocols follow the successful Methods in Molecular Biology series format, each one offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and highly practical, Oligonucleotide Synthesis: Methods and Applications offers today’s investigators not only insight into important new biotechnologies, but also a full range of the detailed protocols needed to work successfully in DNA synthesis and its applications today.