Multiplex Detection of Hotspot Mutations by Rolling Circle-Enabled Universal Microarrays
| Autor: | Gemma Tarafa, Todd Strugnell, Daniela Ladner, John H. Leamon, Stefan Hamann, Deborah A. Dillon, Jose Costa, Paul M. Lizardi |
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| Rok vydání: | 2001 |
| Předmět: |
Microarray
Biology Polymerase Chain Reaction Sensitivity and Specificity Pathology and Forensic Medicine law.invention Automation chemistry.chemical_compound law Neoplasms Multiplex polymerase chain reaction Tumor Cells Cultured Humans Multiplex Molecular Biology Polymerase chain reaction Oligonucleotide Array Sequence Analysis Genetics DNA Neoplasm Cell Biology chemistry Nat Rolling circle replication Mutation DNA microarray DNA |
| Zdroj: | Scopus-Elsevier |
| ISSN: | 0023-6837 |
| Popis: | Detection of somatic low abundance mutations in early cancer development requires a discriminatory, specific, and high-throughput methodology. In this study we report specific, discriminatory detection of low abundance mutations through a novel combination of rolling circle amplification (Nat Genet 1998; 19:225-232) and PCR ligation detection reaction on a universal oligonucleotide microarray (J Mol Biol 1999; 292:251-262). After mutation-specific multiplex ligation and hybridization of 17 pairs of probes to a generic microarray, the ligated probes were visualized. The multiplex mutation-specific ligation is possible only because rolling circle amplification permits quantification of previously undetectable hybridization events conducive to the detection of a single mutation from within a pool of over 100 wild-type alleles. This system is readily adaptable to high-throughput automation using a robot such as the Biomek platform. |
| Databáze: | OpenAIRE |
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