title: Biotin-enhanced fragmentation for direct deoxyribonucleic acid sequencing using matrix-assisted laser desorption/ionization mass spectrometry.
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.
authors.affiliation: Life Sciences Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831-6378, USA.
authors.forename: Z
authors.initials: Z
authors.lastname: Liu
authors.forename: N R
authors.initials: NR
authors.lastname: Isola
authors.forename: S L
authors.initials: SL
authors.lastname: Allman
authors.forename: C H
authors.initials: CH
authors.lastname: Chen
mesh_terms: D001483:Base Sequence; D001710:Biotin / Q000737:chemistry*; D004247:DNA / Q000737:chemistry*; D009841:Oligonucleotides / Q000032:analysis; D017422:Sequence Analysis, DNA / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D009841:Oligonucleotides; D001710:Biotin; D004247:DNA