This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Jeffreys, A. J. Articles by May, C. A. Search for Related Content PubMed PubMed Citation Articles by Jeffreys, A. J. Articles by May, C. A. Social Bookmarking                   What's this?

Methods

DNA Enrichment by Allele-Specific Hybridization (DEASH): A Novel Method for Haplotyping and for Detecting Low-Frequency Base Substitutional Variants and Recombinant DNA Molecules

Department of Genetics, University of Leicester, Leicester LE1 7RH, UK

Detecting rare sequence variants in genomic DNA is central to the analysis of de novo mutation and recombination events and the detection of rare pathological mutations in mixed cell populations. Current PCR techniques suffer from noise that limits detection to variants present at a frequency of at least 10^-4-10^-5 per cell. We nowdescribe an alternative approach that recovers genomic DNA molecules containing a known single-nucleotide variant by hybridization selection using a biotinylated allele-specific oligonucleotide, followed by hybrid capture on streptavidin-coated paramagnetic beads and subsequent analysis by PCR. This technique of DNA enrichment by allele-specific hybridization (DEASH) is fast, effective for all tested single-nucleotide polymorphisms (SNPs), and can recover large (>10 kb) single-stranded molecules. A single round of DEASH is effective in separating haplotypes from genomic DNA and can not only readily detect and validate DNA molecules containing a single base change at a frequency of 10^-5 per cell, but can also place these changes within the context of an extended haplotype. This technique offers a newapproach to the analysis of mutation and recombination, and has the potential to detect very rare de novo base substitutions.

Article and publication are at http://www.genome.org/cgi/doi/10.1101/gr.1214603.

CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				A. J. Jeffreys and R. Neumann Factors influencing recombination frequency and distribution in a human meiotic crossover hotspot Hum. Mol. Genet., August 1, 2005; 14(15): 2277 - 2287. [Abstract] [Full Text] [PDF]