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Eur. J. Mass Spectrom.
9, 213–219 (2003)
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| Biotin-enhanced fragmentation for direct deoxyribo nucleic acid sequencing using matrix-assisted laser desorption/ionization mass spectrometry | ||
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Z. Liu, N.R. Isola, S.L. Allman and C.H. Chen* |
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| 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. | ||
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Keywords: matrix-assisted laser desorption/ionization, time-of-flight mass spectrometry, DNA sequencing, biotin |
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