Oligonucleotide techniques

Conceptually the most simple syntheses of complex molecules involve the joining of structural units in which all functional groups and all asymmetric centres are preformed. This technique can usually only be applied to compounds in which these units are connected by —C—X— bonds rather than C—C. It is illustrated here by the standard syntheses of oligonucleotides, peptides, and polydentate macrocyclic ligands. 

The three techniques — laser desorption ionization, laser ablation with secondary ionization, and matrix-assisted laser desorption — are all used for mass spectrometry of a wide variety of substances from rock, ceramics, and bone to proteins, peptides, and oligonucleotides. 

Another technique often used to examine the stmcture of double-heUcal oligonucleotides is two-dimensional nmr spectroscopy (see AfAGNETiC SPIN resonance). This method rehes on measurement of the nuclear Overhauser effects (NOEs) through space to determine the distances between protons (6). The stmcture of an oligonucleotide may be determined theoretically from a set of iaterproton distances. As a result of the complexities of the experiment and data analysis, the quality of the stmctural information obtained is debated. However, nmr spectroscopy does provide information pertaining to the stmcture of DNA ia solution and can serve as a complement to the stmctural information provided by crystallographic analysis. 

While many diseases have long been known to result from alterations in an individual s DNA, tools for the detection of genetic mutations have only recently become widely available. These techniques rely upon the catalytic efficiency and specificity of enzyme catalysts. For example, the polymerase chain reaction (PCR) relies upon the ability of enzymes to serve as catalytic amplifiers to analyze the DNA present in biologic and forensic samples. In the PCR technique, a thermostable DNA polymerase, directed by appropriate oligonucleotide primers, produces thousands of copies of a sample of DNA that was present initially at levels too low for direct detection. 

This spectroscopy has been used as a sensitive and effective analytical technique for the characterization of specific ligand-oligonucleotide duplex non-covalent complexes [138], due to its specificity, sensitivity and quickness as well as its advantage in determining binding stoichiometry. 

Synthetic oligonucleotides are very important tools in the study and manipulation of DNA, including such techniques as site-directed mutagenesis and DNA amplification by the polymerase chain reaction. The techniques for chemical synthesis of oligonucleotides are highly developed. Very efficient automated methodologies based on solid phase synthesis are used extensively in fields that depend on the availability of defined DNA sequences.52... 

Over the past decade the techniques of combinatorial synthesis have received much attention. Solid phase synthesis of polypeptides and oligonucleotides are especially adaptable to combinatorial synthesis, but the method is not limited to these fields. The goal of combinatorial synthesis is to prepare a large number of related... 

Palchetti I, Mascini M (2005) Electrochemical Adsorption Technique for Immobilization of Single-Stranded Oligonucleotides onto Carbon Screen-Printed Electrodes. 261 27-43 Pascal R, Boiteau L, Commeyras A (2005) From the Prebiotic Synthesis of a-Amino Acids Towards a Primitive Translation Apparatus for the Synthesis of Peptides. 259 69-122 Paulo A, see Santos I (2005) 252 45-84 Perez EM, see Leigh DA (2006) 265 185-208 Perret F, see Coleman AW (2007) 277 31-88 Perron H, see Coleman AW (2007) 277 31-88 Pianowski Z, see Winssinger N (2007) 278 311-342 Piestert F, see Gansauer A (2007) 279 25-52... 

Lebaron, P. Catala, P. Fajon, C. Joux, F. Baudart, J. Bernard, L. A new sensitive, whole-cell hybridization technique for detection of bacteria involving a biotinylated oligonucleotide probe targeting rRNA and tyramide signal amplification. Appl. Environ. Microbiol. 1997, 63, 3274-3278. 

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