Sequence determination, nucleic acid base

Cambridge) the determination of base sequences in nucleic acids. 

Fiskin, A. M., and M. Beer. 1965. Determination of Base Sequence in Nucleic Acids with the Electron Microscope. IV. Nucleoside Complexes with Certain Metal Ions. Biochem. 4,1287. 

In the 1970s he developed a widely used technique of using gel electrophoresis to read nucleotide sequences of DNA segments. The same method was developed independently by Frederick Sanger, and they both won the Nobel Prize in chemistry in 1980 for their contributions concerning the determination of base sequences in nucleic acids. ... 

Walter Gilbert, Frederick Sanger For their contributions concerning the determination of base sequences in nucleic acids. ... 

Figure 20. A diagram illu.< trating phylogenetic relationships between the three kingdoms of organisms. The data used to determine these come from the base sequences of nucleic acids. The tree presently has no root because u f have no unambiguous kmtwledge of the ancestral forms (from Engel and Macko, 1993).

Tandem mass spectrometry has become an important tool for determining the sequence of amino acids in protonated peptides98 and the sequence of bases in deprotonated nucleic acids such as DNA.99 Despite the importance and widespread use of CID-MS to sequence peptides and nucleic acids, the mechanistic details of the dissociation processes are poorly understood. A better understanding of the... 

For most of the history of mankind, unraveling the nucleotide sequence of even a quite small nucleic acid was a formidable undertaking. Following 7 years of labor, Robert Holley solved the first such structure, that for an alanine tRNA from yeast, in 1961. This molecule contains a linear chain of 76 nucleotides and includes some unusual bases, which actually help in base sequence determination. For this achievement, Holley shared the Nobel Prize in Physiology or Medicine in 1968. 

The principles of attachment of molecules to polysaccharides with concomitant insolubilization, discussed in the preceding two Sections, also apply to nucleic acids. The insolubilization of nucleic acids and polynucleotides provides materials useful (a) for fractionation and purification of other nucleic acids and related compounds,(b) for multiplication and isolation of single nucleic acid strands by base-pairing, " (c) for base-sequence determination, (d) as afiR-nants, " templates, and substrates for nucleic acid-related enzymes, and (e) as aflBnants for nucleic acid-binding proteins. ... 

We have already seen that the primary structure of a protein determines its secondary and tertiary structures. The same is true of nucleic acids the nature and order of monomer units determine the properties of the whole molecule. Base pairing in both RNA and DNA depends on a series of complementary bases, whether these bases are on different polynucleotide strands, as in DNA, or on the same strand, as is frequently the case in RNA. Sequencing of nucleic acids is now fairly routine, and this relative ease would have amazed the scientists of the 1950s and 1960s. 

Sequence determination should be simple given that there are only four base types per nucleic acid. Because each nucleotide has a mass of around 310 Da (289-329 for DNA, 305-345 for RNA amino acid residues average around 120 Da) a segment of a nucleic acid with a given mass will have only about 40% the number of nucleotides as there would be of amino acids in an equal mass of protein. Therefore,... 

This assay system developed by Chaires [136] is a new, powerful and effective tool based on the fundamental thermodynamic principle of equilibrium dialysis for the discovery of ligands that bind to nucleic acids with structural and sequence selectivity. Here, identical concentrations of various nucleic acid samples are dialysed in dispodialysers against a common ligand solution. At equilibrium, the contents of the ligand bound to each nucleic acid are determined and this is correlated directly to the ligand s specificity to a particular sequence. 

The discovery of the base-paired, double-helical structure of deoxyribonucleic acid (DNA) provides the theoretic framework for determining how the information coded into DNA sequences is replicated and how these sequences direct the synthesis of ribonucleic acid (RNA) and proteins. Already clinical medicine has taken advantage of many of these discoveries, and the future promises much more. For example, the biochemistry of the nucleic acids is central to an understanding of virus-induced diseases, the immune re-sponse, the mechanism of action of drugs and antibiotics, and the spectrum of inherited diseases. 

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