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PLoS One. 2012;7(2):e31597. doi: 10.1371/journal.pone.0031597. Epub 2012 Feb 15.

Winnowing DNA for rare sequences: highly specific sequence and methylation based enrichment.

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1
Department of Physics and Astronomy, University of British Columbia, Vancouver, British Columbia, Canada.

Abstract

Rare mutations in cell populations are known to be hallmarks of many diseases and cancers. Similarly, differential DNA methylation patterns arise in rare cell populations with diagnostic potential such as fetal cells circulating in maternal blood. Unfortunately, the frequency of alleles with diagnostic potential, relative to wild-type background sequence, is often well below the frequency of errors in currently available methods for sequence analysis, including very high throughput DNA sequencing. We demonstrate a DNA preparation and purification method that through non-linear electrophoretic separation in media containing oligonucleotide probes, achieves 10,000 fold enrichment of target DNA with single nucleotide specificity, and 100 fold enrichment of unmodified methylated DNA differing from the background by the methylation of a single cytosine residue.

PMID:
22355378
PMCID:
PMC3280224
DOI:
10.1371/journal.pone.0031597
[Indexed for MEDLINE]
Free PMC Article
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