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DNA Sequencing by Delayed Extraction-Matrix-Assisted Laser Desorption/Ionizaton Time of Flight Mass Spectrometry
Mark T. Roskey, Peter Juhasz, Igor P. Smirnov, Edward J. Takach, Stephen A. Martin and Lawrence A. Haff
Proceedings of the National Academy of Sciences of the United States of America
Vol. 93, No. 10 (May 14, 1996), pp. 4724-4729
Published by: National Academy of Sciences
Stable URL: http://www.jstor.org/stable/38843
Page Count: 6
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Matrix-assisted laser desorption/ionization (MALDI) time of flight mass spectrometry was used to detect and order DNA fragments generated by Sanger dideoxy cycle sequencing. This was accomplished by improving the sensitivity and resolution of the MALDI method using a delayed ion extraction technique (DE-MALDI). The cycle sequencing chemistry was optimized to produce as much as 100 fmol of each specific dideoxy terminated fragment, generated from extension of a 13-base primer annealed on 40- and 50-base templates. Analysis of the resultant sequencing mixture by DE-MALDI identified the appropriate termination products. The technique provides a new non-gel-based method to sequence DNA which may ultimately have considerable speed advantages over traditional methodologies.
Proceedings of the National Academy of Sciences of the United States of America © 1996 National Academy of Sciences