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Lab Invest. 2001 Aug;81(8):1079-86.

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

Author information

  • 1Department of Pathology, Yale New Haven Hospital, Yale University, New Haven, Connecticut, USA.

Erratum in

  • Lab Invest 2001 Oct;81(10):1338.

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.

PMID:
11502859
[PubMed - indexed for MEDLINE]
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