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Methods Mol Biol. 2011;772:179-91. doi: 10.1007/978-1-61779-228-1_10.

DNA microarray-based mutation discovery and genotyping.

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1
Department of Biology and Center for Genomics and Systems Biology, New York University, New York, NY, USA. dgresham@nyu.edu

Abstract

DNA microarrays provide an efficient means of identifying single-nucleotide polymorphisms (SNPs) in DNA samples and characterizing their frequencies in individual and mixed samples. We have studied the parameters that determine the sensitivity of DNA probes to SNPs and found that the melting temperature (T (m)) of the probe is the primary determinant of probe sensitivity. An isothermal-melting temperature DNA microarray design, in which the T (m) of all probes is tightly distributed, can be implemented by varying the length of DNA probes within a single DNA microarray. I describe guidelines for designing isothermal-melting temperature DNA microarrays and protocols for labeling and hybridizing DNA samples to DNA microarrays for SNP discovery, genotyping, and quantitative determination of allele frequencies in mixed samples.

PMID:
22065438
DOI:
10.1007/978-1-61779-228-1_10
[Indexed for MEDLINE]
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