OLIGONUCLEOTIDE SEQUENCING BY FRAGMENTATION IN MATRIX-ASSISTED LASER DESORPTION IONIZATION TIME-OF-FLIGHT MASS-SPECTROMETRY/

Oligonucleotide sequencing by fragmentation in matrix-assisted laser d esorption/ionization time-of-flight (MALDI-TOF) mass spectrometry has been demonstrated, With this sequencing method, the preparation of oli gonucleotide ladder products and gel electrophoresis are not required, thus the sequencing speed potentially can be increased, Sequence info rmation for single strand oligonucleotide segments up to 35 mer was ob tained from fragmentation patterns using a mixture of anthranilic acid and ammonium citrate or 2,4,6-trihydroxyacetophenone and ammonium cit rate as matrices, This method shows that the cleavage of P-O bond at 5 '-linkage and C-O bond at 3'-linkage could be established in the MALDI process. Such cleavages at specific sites in the backbone of oligonuc leotide can provide the foundation for fast oligonucleotide sequencing . (C) 1997 by John Wiley & Sons, Ltd.