Determination of nucleotide sequence

In another series of experiments, a novel approach to the determination of nucleotide sequence was adopted by A. S. Jones, Stacey, and their co-workers. For example, when calf thymus DNA was treated with mercaptoacetic acid in the presence of zinc chloride and anhydrous sodium sulfate, it yielded aldehydo-apurinic acid bis(carboxymethyl) dithioacetal. When degraded with dilute alkali, this afforded dialyzable fragments, which were separated into at least 20 components. Some were identified, including mono-, di-, and tri-nucleotides, thereby revealing that DNA contain regions of at least three linked pyrimidine nucleotides. The same procedure was applied to the DNA isolated from M. phlei ... 

F. Sanger. 1981. Determination of nucleotide sequences in YiSiA Science 214 1205-1210. (PubMed)... 

The exploration of evolution can also be a laboratory science. In favorable cases, PCR amplification of well-preserved samples allows the determination of nucleotide sequences from extinct organisms. Sequences so determined can help authenticate parts of an evolutionary tree con-... 

Sanger, Frederick (1980). Determination of Nucleotide Sequences in DNA. Nobel e-Museum. Available from . 

This phenomenon can be an interesting one if the cleavage specificity is exhibited not only in homopolyribonucleotides but also in heteropolyribonucleotides hke RNA. If so, this kind of metal-catalyzed reaction could be used in the determination of nucleotide sequence. Consequently, we performed some studies to see whether cleavage specificity could also be obtained with RNA. 

Vidgren, G., Palva, 1., Pakkanen, R., Lounatmaa, K., Palva, A., 1992. S-layer protein gene of Lactobacillus brevis cloning by polymerase chain reaction and determination of nucleotide sequence. J. Bacteriol. 174, 7419—7427. 

With both single- and double-surface techniques detection of single-base mismatches (point mutations) is possible. At present principles of the electrochemical detection of the DNA hybridization event and determination of various genomic nucleotide sequences, after the PCR amplification of DNA, are known. On the other hand, a lot of work has to be done to develop and optimize inexpensive and simple electrochemical DNA sensors with performance comparable to that of DNA arrays based on optical detection. Determination of nucleotide sequences without DNA amplification represents a great challenge. To reach this goal it will be necessary to... 

The Determination of Nucleotide Sequences In DNA and RNA.—DNA. The methods developed by Sanger and co-workers, reported in the last volume, which use specific chain-terminators (dideoxyribonucleoside triphosphates) have proved particularly valuable when applied to single-stranded DNA. However, the scope of the method was limited to naturally occurring single-stranded DNA or to duplex DNA in which strands could be readily separated. 

RNA. There have been several further advances in the methodology for determination of nucleotide sequences in RNA. Nearly all of these methods involve addition of terminal radiolabel to RNA in vitro, degradation of RNA, and analysis of labelled degraded RNA by electrophoresis through gels of polyacrylamide. Consequently the methods appear superficially similar to those reported earlier and some do indeed supplement existing methodologies. For example, nucleases... 

The Determination of Nucleotide Sequences in DNA and RNA.—DNA. By electro phoresis on polyacrylamide gels it is possible to resolve DNA strands into bands... 

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