Detection of microarray-hybridized oligonucleotides with magnetic beads

The efficiency of hybridization analysis with oligonucleotide microarrays depends heavily on the method of detection. Conventional methods based on labeling nucleic acids with fluorescent, chemiluminescent, enzyme, or radioactive reporters suffer from a number of serious drawbacks which demand devel...

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Bibliographic Details
Published in:	Analytical biochemistry Vol. 399; no. 1; pp. 125 - 131
Main Authors:	Shlyapnikov, Yury M., Shlyapnikova, Elena A., Morozova, Tamara Ya, Beletsky, Igor P., Morozov, Victor N.
Format:	Journal Article
Language:	English
Published:	United States Elsevier Inc 01-04-2010
Subjects:	Biotin - chemistry DNA microarray DNA, Plant - analysis DNA, Plant - chemistry Glycine max - genetics Hybridization detection Magnetic beads Magnetics Oligonucleotide Array Sequence Analysis - methods Plants, Genetically Modified Streptavidin - chemistry DNA microarray Magnetic beads Hybridization detection
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Summary:	The efficiency of hybridization analysis with oligonucleotide microarrays depends heavily on the method of detection. Conventional methods based on labeling nucleic acids with fluorescent, chemiluminescent, enzyme, or radioactive reporters suffer from a number of serious drawbacks which demand development of new detection techniques. Here, we report two new approaches for detection of hybridization with oligonucleotide microarrays employing magnetic beads as active labels. In the first method streptavidin-coated magnetic beads are used to discover biotin-labeled DNA molecules hybridized with arrayed oligonucleotide probes. In the second method biotin-labeled DNA molecules are bound first to the surface of magnetic beads and then hybridized with arrayed complementary strands on bead-array contacts. Using a simple low-power microscope with a dark-field illumination and a pair of complementary primers as a model hybridization system we evaluated sensitivity, speed, and cost of the new detection method and compared its performance with the detection techniques employing enzyme and fluorescent labels. It was shown that the detection of microarray-hybridized DNA with magnetic beads combines low cost with high speed and enhanced assay sensitivity, opening a new way to routine hybridization assays which do not require precise measurements of DNA concentration.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0003-2697 1096-0309
DOI:	10.1016/j.ab.2009.12.030