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Microbead expression arrays
Genome Biology volume 1, Article number: spotlight-20000606-01 (2000)
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.
Serial analysis of gene expression.
Quantitative monitoring of gene expression patterns with a complementary DNA microarray.
Nature Biotechnology, [http://www.nature.com/nbt/]
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Wells, W. Microbead expression arrays. Genome Biol 1, spotlight-20000606-01 (2000). https://doi.org/10.1186/gb-spotlight-20000606-01
- Restriction Endonuclease
- Confocal Microscopy
- cDNA Sequence
- Cleavage Site
- Fluorescent Probe