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A Strategy for Examining Complex Mixtures of Deoxyoligonucleotides Using IP-RP HPLC, MALDI-TOF MD, and Informatics

Published

Author(s)

Peter Vallone, J E. Devaney, J M. Marino

Abstract

Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS)and ion pair-reverse phase high performance liquid chromatography (IP-RP HPLC) techniques were combined to determine the sequence identity of short single-stranded deoxyoligonucleotides. In demonstrating this methodology a commercially abailable multiplex set of 8 primer pairs used for amplifying 8 specific loci, but with unknown sequences were separated and collected by IP-RP HPLC. Partial sequence information for IP-RP HPLC fractions was obtained from analyzing exonuclease digestion products by MALDI-TOF MS. By utilizing IP-RP HPLC, MALDI-TOF MS, exonuclease digests, and a simple computational algorithm an integrated strategy for determining the sequence of short necleic acid oligomers is demonstrated.
Citation
Analytical Biochemistry
Volume
304

Keywords

informatics, MALDI-TOF MS, nucleic acid, PCR

Citation

Vallone, P. , Devaney, J. and Marino, J. (2002), A Strategy for Examining Complex Mixtures of Deoxyoligonucleotides Using IP-RP HPLC, MALDI-TOF MD, and Informatics, Analytical Biochemistry (Accessed April 19, 2024)
Created March 16, 2002, Updated February 19, 2017