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Selective Circular Oligonucleotide Probes Improve Detection of Point Mutations in DNA
Author(s) -
Tennilä Tuula,
Ketomäki Kaisa,
Penttinen Piia,
Tengvall Unni,
Azhayeva Elena,
Auriola Seppo,
Lönnberg Harri,
Azhayev Alex
Publication year - 2004
Publication title -
chemistry and biodiversity
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.427
H-Index - 70
eISSN - 1612-1880
pISSN - 1612-1872
DOI - 10.1002/cbdv.200490052
Subject(s) - oligonucleotide , chemistry , phosphodiester bond , dna , circular dichroism , electrospray ionization , mass spectrometry , rolling circle replication , combinatorial chemistry , stereochemistry , chromatography , biochemistry , polymerase , gene , rna
The synthesis of disulfide‐cross‐linked circular oligonucleotides, employing two different approaches, was accomplished. Several circular oligomers, which bear a C (5)‐aminoalkyl‐tethered thymidine unit, were labeled with photoluminescent europium(III) chelates. All circular structures were thoroughly characterized with denaturing PAGE and electrospray‐ionization mass spectrometry. It was demonstrated that the disulfide cross‐linking, resulting in circularization, considerably increases the enzymatic stability of phosphodiester oligonucleotides. In addition, UV melting experiments, followed, where possible, by extraction of thermodynamic parameters, revealed that several circular oligomers appear to be more selective towards their complementary targets than their corresponding linear precursors. Finally, the mixed‐phase hybridization experiments have demonstrated that use of circular probes indeed improves the selectivity in the detection of DNA point mutations.