Braslavsky, Ido and Hebert, Benedict and Kartalov, Emil and Quake, Stephen R. (2003) Sequence information can be obtained from single DNA molecules. Proceedings of the National Academy of Sciences of the United States of America, 100 (7). pp. 3960-3964. ISSN 0027-8424. http://resolver.caltech.edu/CaltechAUTHORS:BRApnas03
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The completion of the human genome draft has taken several years and is only the beginning of a period in which large amounts of DNA and RNA sequence information will be required from many individuals and species. Conventional sequencing technology has limitations in cost, speed, and sensitivity, with the result that the demand for sequence information far outstrips current capacity. There have been several proposals to address these issues by developing the ability to sequence single DNA molecules, but none have been experimentally demonstrated. Here we report the use of DNA polymerase to obtain sequence information from single DNA molecules by using fluorescence microscopy. We monitored repeated incorporation of fluorescently labeled nucleotides into individual DNA strands with single base resolution, allowing the determination of sequence fingerprints up to 5 bp in length. These experiments show that one can study the activity of DNA polymerase at the single molecule level with single base resolution and a high degree of parallelization, thus providing the foundation for a practical single molecule sequencing technology.
|Additional Information:||© 2003 by The National Academy of Sciences. Communicated by Harry B. Gray, California Institute of Technology, Pasadena, CA, January 25, 2003 (received for review November 18, 2002). We thank Henry Lester and Marc Unger for fruitful discussions. Partial financial support was provided by the Lester Deutsch fellowship, National Institutes of Health Grant HG01641-01, the Packard Foundation, and the Burroughs–Wellcome Foundation.|
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|Deposited On:||14 Sep 2005|
|Last Modified:||14 Nov 2014 19:18|
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