DNA Sequencing by Delayed Extraction-Matrix-Assisted Laser Desorption/Ionizaton Time of Flight Mass Spectrometry

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...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1996-05, Vol.93 (10), p.4724-4729
Main Authors: Roskey, Mark T., Juhasz, Peter, Smirnov, Igor P., Takach, Edward J., Martin, Stephen A., Haff, Lawrence A.
Format: Article
Language:English
Subjects:
Biochemistry
Dimers
DNA
Gels
Ions
Lasers
Mass spectra
Mass spectroscopy
Oligonucleotides
Sequencing
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Summary: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.
ISSN:0027-8424
DOI:10.1073/pnas.93.10.4724