Analysis of negatively 'charge tagged' DNA by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.
Rapid Commun Mass Spectrom. 1999; 13(17):1739-43.RC

Abstract

An improvement in detectability and stability of DNA analysis by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) using oligonucleotides modified with a neutralized backbone and a fixed single, positive charge was recently reported. The attachment of the positive charge requires a primary amino group, limiting this approach to accordingly functionalized DNA. The method described here uses backbone neutralized DNA for the same purpose, with a single unmodified phosphate in the DNA backbone carrying the negative charge. Thus, no chemical modification other than neutralizing the remaining charges on the phosphorothioates is required. This is performed in a single methylation step. The enhancement in sensitivity is comparable to that for DNA carrying a single positive charge, interestingly even when using the same non-protonating matrix. The mechanistic implications of these findings regarding the MALDI process are discussed. The DNA derivatization methods presented help to make MALDI-MS of DNA applicable and competitive for genome analysis and medical diagnostics.

Authors+Show Affiliations

Berlin K

Max-Planck-Institute for Molecular Genetics, Department Lehrach, Ihnestrasse 73, 14195 Berlin, Germany.

Gut IG

No affiliation info available

MeSH

DNAMolecular StructureOligonucleotidesPhosphatesSpectrometry, Mass, Matrix-Assisted Laser Desorption-IonizationStatic Electricity

Pub Type(s)

Journal Article

Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

10455243

Citation

Berlin, K, and I G. Gut. "Analysis of Negatively 'charge Tagged' DNA By Matrix-assisted Laser Desorption/ionization Time-of-flight Mass Spectrometry." Rapid Communications in Mass Spectrometry : RCM, vol. 13, no. 17, 1999, pp. 1739-43.

Berlin K, Gut IG. Analysis of negatively 'charge tagged' DNA by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Rapid Commun Mass Spectrom. 1999;13(17):1739-43.

Berlin, K., & Gut, I. G. (1999). Analysis of negatively 'charge tagged' DNA by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Rapid Communications in Mass Spectrometry : RCM, 13(17), 1739-43.

Berlin K, Gut IG. Analysis of Negatively 'charge Tagged' DNA By Matrix-assisted Laser Desorption/ionization Time-of-flight Mass Spectrometry. Rapid Commun Mass Spectrom. 1999;13(17):1739-43. PubMed PMID: 10455243.

* Article titles in AMA citation format should be in sentence-case

TY - JOUR T1 - Analysis of negatively 'charge tagged' DNA by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. AU - Berlin,K, AU - Gut,I G, PY - 1999/8/24/pubmed PY - 1999/8/24/medline PY - 1999/8/24/entrez SP - 1739 EP - 43 JF - Rapid communications in mass spectrometry : RCM JO - Rapid Commun Mass Spectrom VL - 13 IS - 17 N2 - An improvement in detectability and stability of DNA analysis by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) using oligonucleotides modified with a neutralized backbone and a fixed single, positive charge was recently reported. The attachment of the positive charge requires a primary amino group, limiting this approach to accordingly functionalized DNA. The method described here uses backbone neutralized DNA for the same purpose, with a single unmodified phosphate in the DNA backbone carrying the negative charge. Thus, no chemical modification other than neutralizing the remaining charges on the phosphorothioates is required. This is performed in a single methylation step. The enhancement in sensitivity is comparable to that for DNA carrying a single positive charge, interestingly even when using the same non-protonating matrix. The mechanistic implications of these findings regarding the MALDI process are discussed. The DNA derivatization methods presented help to make MALDI-MS of DNA applicable and competitive for genome analysis and medical diagnostics. SN - 0951-4198 UR - https://www.unboundmedicine.com/medline/citation/10455243/Analysis_of_negatively_'charge_tagged'_DNA_by_matrix_assisted_laser_desorption/ionization_time_of_flight_mass_spectrometry_ L2 - https://doi.org/10.1002/(SICI)1097-0231(19990915)13:17<1739::AID-RCM708>3.0.CO;2-7 DB - PRIME DP - Unbound Medicine ER -

Tags

Analysis of negatively 'charge tagged' DNA by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.Rapid Commun Mass Spectrom. 1999; 13(17):1739-43.RC