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

被引：1
作者：
Liu, Z [1 ]
Isola, NR [1 ]
Allman, SL [1 ]
Chen, CH [1 ]
机构：
[1] Oak Ridge Natl Lab, Div Life Sci, Oak Ridge, TN 37831 USA
来源：
EUROPEAN JOURNAL OF MASS SPECTROMETRY | 2003年 / 9卷 / 03期
关键词：
matrix-assisted laser desorption/ionization; time-of-flight mass spectrometry; DNA sequencing; biotin;
D O I：
10.1255/ejms.542
中图分类号：
O64 [物理化学（理论化学）、化学物理学]; O56 [分子物理学、原子物理学];
学科分类号：
070203 ; 070304 ; 081704 ; 1406 ;
摘要：
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 fact that 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.
引用
收藏
页码：213 / 219
页数：7
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