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.2001 Aug;81(8):1079-86.
doi: 10.1038/labinvest.3780320.

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

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

D P Ladner et al. Lab Invest.2001 Aug.
Free article

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.

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