Synthesis and evaluation of through bond energy transfer flourescent dyes and new terminators for DNA sequencing /
A set of four through-bond fluorescence energy crographics.
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| Format: | Thesis Book |
| Language: | English |
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[Place of publication not identified] :
[publisher not identified] ;
1998.
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| Summary: | A set of four through-bond fluorescence energy crographics. transfer (ET) cassettes has been prepared and the fluorescence properties examined. These cassettes were made from four BODIPY dyes with distinct emission maxima. Efficient ET was observed in all four dyes. ET enhancements were up to 12 times that of the single dye. However, the two 3,5-diaryl substituted BODIPYS 14 and 15 exhibited more than one order of less intense fluorescence than the alkyl substituted BODIPYV 12 and 13. As a result, the two long wavelength ET dye cassettes 20 and 21 were also more than one order of magnitude less intense than the two short wavelength cassettes making the four, as a set, unsuitable for four-color fluorescence detected DNA sequencing. Two more ET cassettes were prepared having anthracene or perylene as the donor and BODIPYO 13 [(Xmas = 529 nm, km = 543 nm)] as the common acceptor. These two ET cassettes (AET and PET) were compared with ET cassette BET-; which had the same acceptor. Of the three, BET-Z, having BODIPY* 12 [(lmax = 504 nm, 'em = 515nm)] as the donor, had the most intense emission from the acceptor when the ET cassettes were excited at 488 nm or 514 nm. When excited at the respective absorption maxima of each donor, PET and BET-; had very similar emission intensities. The AET system, having anthracene as the donor, had approximately 25% the intensity of PET and BET-2 when each was excited at the absorption maximum of the donor. Two new types of chain terminators for DNA sequencing have also been based dideoxynucleoside triphosphate (ddNTP) analogues containing 3-nitropyrrole or 5-nitroindole as the heterocycle were not incorporated by any of the DNA polymerizes tested. However, a rigid carboacyclic adenine nucleotide triphosphate was found to be an efficient terminator of template dependent DNA polymerization. This triphosphate was incorporated at concentrations comparable to those at which ddNTPs are used in sequencing. Comparison of [Kau/vm] values shows that the analogue is incorporated 48 times less efficiently than ddATP by AmpliTaq* FS DNA polymerase. |
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| Item Description: | "Major subject: Chemistry". Vita. |
| Physical Description: | xi, 147 leaves : illustrations ; 28 cm. |
| Bibliography: | Includes bibliographical references: pages 44-46. |