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Multiplexed hybridizations of positively charge‐tagged peptide nucleic acids detected by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry
Author(s) -
Bauer Oliver,
Guerasimova Anna,
Sauer Sascha,
Thamm Sabine,
Steinfath Matthias,
Herwig Ralf,
Janitz Michal,
Lehrach Hans,
Radelof Uwe
Publication year - 2004
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.1554
Subject(s) - chemistry , oligonucleotide , nucleic acid , complementary dna , peptide nucleic acid , mass spectrometry , dna , genomic dna , nucleic acid thermodynamics , chromatography , combinatorial chemistry , biochemistry , gene , base sequence
Peptide nucleic acid (PNA) is a novel class of DNA analogues in which the entire sugar‐phosphate backbone is replaced by a pseudopeptide counterpart. Owing to its neutral character and the consequent lack of electrostatic repulsion, PNA exhibits very stable heteroduplex formation with complementary nucleic acid that is essentially ionic strength independent and enables hybridization under minimum salt conditions. This feature as well as its superior ion stability and easy ionization compared to DNA renders PNA very attractive for hybridization‐based matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOFMS) applications. We have developed an approach to DNA characterization that takes advantage of multiplexed PNA hybridizations analyzed by MALDI‐TOFMS. Our motivation was the further development of oligonucleotide fingerprinting, an efficient technique for cDNA and genomic DNA library characterization. Through positive ‘charge‐tagging’ of PNA the efficiency of detection in MALDI‐TOFMS was considerably enhanced permitting an unparalleled degree of multiplexing. Results from the simultaneous hybridization of 21 charge‐tagged PNA hexamer oligonucleotides showed that genomic DNA and cDNA clones are successfully characterized on the basis of their hybridization profiles. The degree of multiplexing achieved may render a significant increase in throughput and hence efficiency of oligonucleotide fingerprinting possible. Copyright © 2004 John Wiley & Sons, Ltd.