Nucleobase analogs for degenerate hybridization devised through conformational pairing analysis
Autor: | Michelle L. Abraham, Maria Albalos, Toumy Guettouche, Michel J. Friesenhahn, Thomas R. Battersby |
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Jazyk: | angličtina |
Rok vydání: | 2007 |
Předmět: | |
Zdroj: | BioTechniques, Vol 43, Iss 5, Pp 617-624 (2007) |
Druh dokumentu: | article |
ISSN: | 1940-9818 0736-6205 |
DOI: | 10.2144/000112602 |
Popis: | A conformational pairing analysis was used to devise nucleobase analogs capable of forming nonselective and energetically favorable base pairs opposite either the purine or the pyrimidine constituents of nucleic acids. 5-methylisocytosine and isoguanine were conceived as a degenerate pyrimidine and a degenerate purine, respectively. Data from previous DNA duplex melting experiments verified that the analogs can act as degenerate nucleobases as hypothesized. Isoguanine also formed unusually stable base pairs with guanine. A quantitative PCR assay yielding equivalent results across hepatitis C virus (HCV) subtypes was created with this system, despite the use of a single probe targeted to a polymorphic region. Amplification curves using probes with 5-methylisocytosine or isoguanine opposite appropriate ambiguous target positions exhibited more signal than curves from similar probes containing common degenerate nucleobase hypoxanthine. |
Databáze: | Directory of Open Access Journals |
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