Use of peptide nucleic acid probes for detecting DNA single-base mutations by capillary electrophoresis
Articolo
Data di Pubblicazione:
2002
Citazione:
Use of peptide nucleic acid probes for detecting DNA single-base mutations by capillary electrophoresis / A. Basile, A. Giuliani, G. Pirri, M. Chiari. - In: ELECTROPHORESIS. - ISSN 0173-0835. - 23:6(2002 Mar), pp. 926-929.
Abstract:
Peptide nucleic acid (PNA) oligomers can be used as probes in pre-gel hybridization experiments, as an alternative to Southern hybridization. In this technique, the PNA probe is hybridized to a cyanine-5 labeled DNA sample denatured at low ionic strength, and the mixture is directly injected for size separation into a capillary electrophoresis (CE) system equipped with laser-induced fluorescence (LIF) detector. The neutral backbone of PNA allows hybridization to occur at low ionic strength and assures an efficient CE separation of the PNA/DNA hybrids from both double-stranded and single-stranded DNA. We have used as a model system the cystic fibrosis R553X and R1162X single-base mutations and we have assessed the influence of various factors, such as temperature and denaturants concentration on DNA/PNA hybrid stability in order to achieve the high specificity required for a single base pair discrimination.
Tipologia IRIS:
01 - Articolo su periodico
Keywords:
Capillary electrophoresis; Hybridization; Mutation detection; Nucleic acid; Peptide nucleic acid
Elenco autori:
A. Basile, A. Giuliani, G. Pirri, M. Chiari
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