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Proceedings Paper

Dye terminator chemistry: effects of substrate structure on sequencing analysis
Author(s): Shaheer H. Khan; Barnett B. Rosenblum; Weiguo Zhen; Meng Taing; Steve Menchen
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Paper Abstract

Dye-labeled dideoxy terminators are the preferred reporter groups for DNA sequencing. This labeling technique can result in uneven peak heights in the electropherograms due to sequence context effects on the biochemical distribution of extension vs termination events. Large variations in peak heights can result in difficulty in identifying very small peaks adjacent to big peaks. Using substrate structure to minimize the sequence context effects on dye-terminator incorporation, we have studied the effects of combinations of base, dye, and dye linkage on terminator incorporation during sequencing. We have identified a propargyl ethoxyamino linker that proved useful in improving peak height evenness and enhancing the terminator activity. New dye-linker-base combinations were optimized for overall peak evenness and relative electrophoretic mobility shifts. The more even peak patterns improve base calling and heterozygote analysis using dye terminator chemistry.

Paper Details

Date Published: 3 May 1999
PDF: 12 pages
Proc. SPIE 3602, Advances in Fluorescence Sensing Technology IV, (3 May 1999); doi: 10.1117/12.347536
Show Author Affiliations
Shaheer H. Khan, Perkin Elmer Corp. (United States)
Barnett B. Rosenblum, Perkin Elmer Corp. (United States)
Weiguo Zhen, Perkin Elmer Corp. (United States)
Meng Taing, Perkin Elmer Corp. (United States)
Steve Menchen, Perkin Elmer Corp. (United States)

Published in SPIE Proceedings Vol. 3602:
Advances in Fluorescence Sensing Technology IV
Joseph R. Lakowicz; Steven A. Soper; Richard B. Thompson, Editor(s)

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