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June 20, 2011
Volume 89, Number 25
p. 32

Fast, Sensitive DNA Sequencing

Combining pyrosequencing and fluorescence measurements leads to a quick and sensitive method for analyzing DNA

Celia Henry Arnaud

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fluorescent

Fluorogenic pyrosequencing, a new DNA sequencing method, combines the speed and one-color detection of conventional pyrosequencing with the sensitivity of fluorescence-based methods (Nat. Methods, DOI: 10.1038/nmeth.1629). In the new method, as in other sequencing-by-synthesis methods, the sequence of a target DNA molecule is determined by identifying the order in which nucleotides are incorporated when that DNA is used as a template for DNA synthesis. X. Sunney Xie and coworkers at Harvard University label all four nucleotides with an identical dye, which is nonfluorescent when it is attached to a nucleotide. Adding a nucleotide to the growing DNA strand releases the dye, which becomes fluorescent. The dye is trapped in polymeric microreactors, in which the DNA is tethered. The fluorescence is detected and the dye is washed away. Repeating the cycle many times with each of the four nucleotides reveals the sequence of the DNA template. Xie and coworkers believe the method will offer low cost, high throughput, and rapid turnaround.

Chemical & Engineering News
ISSN 0009-2347
Copyright © 2011 American Chemical Society
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