Use of a PNA probe to block DNA-mediated PCR product formation in prokaryotic RT-PCR
A novel method eliminating DNA-mediated PCR product formation in reverse transcription PCR (RT-PCR) amplification of specific RNA sequences is described. The method exploits the higher melting temperature values of peptide nucleic acid (PNA)/DNA duplexes compared with DNA/DNA duplexes by binding a s...
| Veröffentlicht in: | BioTechniques. - 1993. - 42(2007), 5 vom: 01. Mai, Seite 609-10, 612-4 |
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| Weitere Verfasser: | , , |
| Format: | Aufsatz |
| Sprache: | English |
| Veröffentlicht: |
2007
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| Zugriff auf das übergeordnete Werk: | BioTechniques |
| Schlagworte: | Journal Article Research Support, Non-U.S. Gov't DNA Probes DNA, Bacterial Nucleic Acid Heteroduplexes Peptide Nucleic Acids DNA 9007-49-2 |
| Zusammenfassung: | A novel method eliminating DNA-mediated PCR product formation in reverse transcription PCR (RT-PCR) amplification of specific RNA sequences is described. The method exploits the higher melting temperature values of peptide nucleic acid (PNA)/DNA duplexes compared with DNA/DNA duplexes by binding a sequence-specific PNA probe to a genomic sequence immediately overlapping one of the PCR-primer attachment sites within the sequence of interest. Hybridization of the blocking probe precludes primer attachment to DNA without affecting attachment of the same primer to the reverse transcription-generated cDNA sequence. A four-step PCR cycle is used that allows the PNA probe to hybridize to the DNA strand at a higher temperature just prior to the primer annealing step |
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| Beschreibung: | Date Completed 22.06.2007 Date Revised 07.06.2018 published: Print Citation Status MEDLINE |
| ISSN: | 0736-6205 |