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Additional info for 21st Century Medicine (Scientific American Special Online Issue No. 30)
The fi nal round yields copy fragments of varying lengths, each terminating with a fluorescently tagged base. Separating these fragments by size in a process called electrophoresis, then reading the fluorescent signal of each terminal tag as it passes by a viewer, provides the sequence of bases in the original strand [see box on preceding two pages]. Reliability and accuracy are advantages of Sanger sequencing, although even with refinements over the years, the method remains time-consuming and expensive.
Grouped under the heading of nanopore sequencing, these methods focus on the physical differences between the four base types to produce a readable signal. 5-nanometer pore, it causes fluctuations in the pore’s electrical conductance. Each base type produces a slightly different conductance change that can be used to identify it [see box above]. Devised by Dan Branton of Harvard, Dave Deamer of the University of California, Santa Cruz, and me, this method is in development now by Agilent Technologies and others with interesting variations, such as fluorescent signal detection.
Jay Shendure, Robi D. Mitra, Chris Varma and George M. Church in Nature Reviews Genetics, Vol. 5, pages 335–344; May 2004. How Sequencing Is Done. S. Dept. of Energy, Office of Science, updated September 9, 2004. html NHGRI Seeks Next Generation of Sequencing Technologies. gov/12513210 Accurate Multiplex Polony Sequencing of an Evolved Bacterial Genome. Jay Shendure et al. in Science, Vol. 309, pages 1728–1732; September 9, 2005. Genome Sequencing in Microfabricated High-Density Picolitre Reactors.
21st Century Medicine (Scientific American Special Online Issue No. 30) by Scientific American