Multiplex detection of hotspot mutations by rolling circle-enabled universal microarrays

Daniela P. Ladner, John H. Leamon, Stefan Hamann, Gemma Tarafa, Todd Strugnell, Deborah Dillon, Paul Lizardi, José Costa*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

19 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)1079-1086
Number of pages8
JournalLaboratory Investigation
Volume81
Issue number8
DOIs
StatePublished - 2001

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Molecular Biology
  • Cell Biology

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