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Biofunctionalization of Polyoxometalates with DNA Primers, Their Use in the Polymerase Chain Reaction (PCR) and Electrochemical Detection of PCR Products
Author(s) -
Debela Ahmed M.,
Ortiz Mayreli,
Beni Valerio,
Thorimbert Serge,
Lesage Denis,
Cole Richard B.,
O'Sullivan Ciara K.,
Hasenknopf Bernold
Publication year - 2015
Publication title -
chemistry – a european journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.687
H-Index - 242
eISSN - 1521-3765
pISSN - 0947-6539
DOI - 10.1002/chem.201502247
Subject(s) - primer (cosmetics) , capillary electrophoresis , bioconjugation , polymerase chain reaction , chemistry , primer dimer , dna , electrochemistry , electrophoresis , primer extension , microbiology and biotechnology , combinatorial chemistry , chromatography , biochemistry , multiplex polymerase chain reaction , biology , electrode , gene , organic chemistry , base sequence
The bioconjugation of polyoxometalates (POMs), which are inorganic metal oxido clusters, to DNA strands to obtain functional labeled DNA primers and their potential use in electrochemical detection have been investigated. Activated monooxoacylated polyoxotungstates [SiW 11 O 39 {Sn(CH 2 ) 2 CO}] 8− and [P 2 W 17 O 61 {Sn(CH 2 ) 2 CO}] 6− have been used to link to a 5′‐NH 2 terminated 21‐mer DNA forward primer through amide coupling. The functionalized primer was characterized by using a battery of techniques, including electrophoresis, mass spectrometry, as well as IR and Raman spectroscopy. The functionality of the POM‐labeled primers was demonstrated through hybridization with a surface‐immobilized probe. Finally, the labeled primers were successfully used in the polymerase chain reaction (PCR) and the PCR products were characterized by using electrophoresis.