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Microbead expression arrays
Strategies for expression analysis range from exhaustive sequencing (and thus counting) of cDNAs to hybridization arrays.
Email: William Wells - wells@biotext.com News from The Scientist 2000, 1(1):20000606-03
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Strategies for expression analysis range from exhaustive sequencing (and thus counting) of cDNAs to hybridization arrays. In the June issue of Nature Biotechnology Brenner et al. describe a method that combines the digital precision of the former with the speed and throughput of the latter (Nat. Biotech. 2000, 18:630-634). Brenner et al. attach tagged cDNAs to microbeads and then sequence the overhanging ends of the cDNAs by detecting the hybridization of fluorescently labeled probes. After one overhang is identified, a binding site for a type IIs restriction endonuclease (within the probe) is used to cleave a distant cleavage site (within the cDNA sequence) to expose a new overhang. The coming and going of fluorescent probes is monitored by confocal microscopy of the microbeads, which are immobilized in a flow cell. Hundreds of thousands of mRNAs are identified in a few days, exceeding the throughput per machine of conventional sequencers by over 10-fold.
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