Zip nucleic acids are potent hydrolysis probes for quantitative PCR
| Autor: | Nathalie Lenne-Samuel, Clément Paris, Patrick Erbacher, Valérie Moreau, Gaëlle Deglane, Emilie Voirin |
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| Rok vydání: | 2010 |
| Předmět: |
chemistry.chemical_classification
Base Pair Mismatch Oligonucleotide Hydrolysis Cationic polymerization Biology Nucleic Acid Denaturation Polymerase Chain Reaction Polymorphism Single Nucleotide Real-time polymerase chain reaction Biochemistry chemistry Molecular beacon Genetics Nucleic acid Methods Online Spermine Nucleotide Oligonucleotide Probes DNA Primers |
| Zdroj: | Nucleic Acids Research |
| ISSN: | 1362-4962 0305-1048 |
| DOI: | 10.1093/nar/gkp1218 |
| Popis: | Zip nucleic acids (ZNAs) are oligonucleotides conjugated with cationic spermine units that increase affinity for their target. ZNAs were recently shown to enable specific and sensitive reactions when used as primers for polymerase chain reaction (PCR) and reverse-transcription. Here, we report their use as quantitative PCR hydrolysis probes. Ultraviolet duplex melting data demonstrate that attachment of cationic residues to the 3' end of an oligonucleotide does not alter its ability to discriminate nucleotides nor the destabilization pattern relative to mismatch location in the oligonucleotide sequence. The stability increase provided by the cationic charges allows the use of short dual-labeled probes that significantly improve single-nucleotide polymorphism genotyping. Longer ZNA probes were shown to display reduced background fluorescence, therefore, generating greater sensitivity and signal level as compared to standard probes. ZNA probes thus provide broad flexibility in assay design and also represent an effective alternative to minor groove binder- and locked nucleic-acid-containing probes. |
| Databáze: | OpenAIRE |
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