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Eur. J. Mass Spectrom. DOI: 10.1255/ejms.542

Biotin-enhanced fragmentation for direct deoxyribo nucleic acid sequencing using matrix-assisted laser desorption/ionization mass spectrometry

Z. Liu, N.R. Isola, S.L. Allman and C.H. Chen*
Life Sciences Division, Oak Ridge National Laboratory, Oak Ridge, TN 37, USA

ABSTRACT:
Fragmentation of synthetic oligonucleotides under the influence of biotin was investigated using 3-hydroxypicolinic acid (3-HPA) as a matrix-assisted laser desorption/ionization (MALDI) matrix. Addition of biotin into the sample enhanced fragmentation of the oligonucleotide between bases. However, when the biotin was tagged to the 5′-terminus of the oligonucleotide, enhancements were observed not only in desorption/ionization efficiency but also in the fragmentation of molecular ions. The protonation/deprotonation process occurs on the tagged biotin is a possible reason for the enhancement in desorption/ionization. Site-specific backbone cleavage fragmentation patterns were observed. The sequences of oligonucleotides can be obtained from their fragment ions. The direct sequencing of a 5′-biotin-tagged 25-mer is demonstrated.

Keywords: matrix-assisted laser desorption/ionization, time-of-flight mass spectrometry, DNA sequencing, biotin


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