Resonance ionization spectroscopy for multiplex sequencing of Tin-labeled DNA

A method is described for synthesis of a tin reagent, triethylstannylpropanoic acid (TESDA), and its attachment to oligonucleotide primers. Except for the expected mobility retardation, the presence of [ 116Sn]-TESDA did not affect the sequencing ladder on electrophoresis gels. By using [ 120Sn]-TES...

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Veröffentlicht in:Genetic analysis, techniques and applications techniques and applications, 1991-09, Vol.8 (6), p.167-170
Hauptverfasser: Sachleben, Richard A., Brown, Gilbert M., Sloop, Frederick V., Arlinghaus, Heinrich F., England, M.Wendy, Foote, Robert S., Larimer, Frank W., Woychik, Richard P., Thonnard, Norbert, Jacobson, K.Bruce
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Sprache:eng
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Zusammenfassung:A method is described for synthesis of a tin reagent, triethylstannylpropanoic acid (TESDA), and its attachment to oligonucleotide primers. Except for the expected mobility retardation, the presence of [ 116Sn]-TESDA did not affect the sequencing ladder on electrophoresis gels. By using [ 120Sn]-TESDA and [ 35S]-dTTP simultaneously in the Sanger procedure, DNA bands on an electrophoresis gel were first located by autoradiography and then by resonance ionization spectroscopy to demonstrate the coincidence of the signals. Previous results using stable isotopes as labels on model compounds are now confirmed by their use in actual DNA sequencing products.
ISSN:1050-3862
DOI:10.1016/1050-3862(91)90057-X