DNA sequencing technology, walking with modular primers. Final report

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The success of the Human Genome Project depends on the development of adequate technology for rapid and inexpensive DNA sequencing, which will also benefit biomedical research in general. The authors are working on DNA technologies that eliminate primer synthesis, the main bottleneck in sequencing by primer walking. They have developed modular primers that are assembled from three 5-mer, 6-mer or 7-mer modules selected from a presynthesized library of as few as 1,000 oligonucleotides ({double_bond}4, {double_bond}5, {double_bond}7). The three modules anneal contiguously at the selected template site and prime there uniquely, even though each is not unique for the most part ... continued below

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4 p.

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Ulanovsky, L. December 31, 1996.

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Description

The success of the Human Genome Project depends on the development of adequate technology for rapid and inexpensive DNA sequencing, which will also benefit biomedical research in general. The authors are working on DNA technologies that eliminate primer synthesis, the main bottleneck in sequencing by primer walking. They have developed modular primers that are assembled from three 5-mer, 6-mer or 7-mer modules selected from a presynthesized library of as few as 1,000 oligonucleotides ({double_bond}4, {double_bond}5, {double_bond}7). The three modules anneal contiguously at the selected template site and prime there uniquely, even though each is not unique for the most part when used alone. This technique is expected to speed up primer walking 30 to 50 fold, and reduce the sequencing cost by a factor of 5 to 15. Time and expensive will be saved on primer synthesis itself and even more so due to closed-loop automation of primer walking, made possible by the instant availability of primers. Apart from saving time and cost, closed-loop automation would also minimize the errors and complications associated with human intervention between the walks. The author has also developed two additional approaches to primer-library based sequencing. One involves a branched structure of modular primers which has a distinctly different mechanism of achieving priming specificity. The other introduces the concept of ``Differential Extension with Nucleotide Subsets`` as an approach increasing priming specificity, priming strength and allowing cycle sequencing. These approaches are expected to be more robust than the original version of the modular primer technique.

Physical Description

4 p.

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OSTI as DE99002793

Medium: P; Size: 4 p.

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  • Other Information: PBD: [1996]

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  • Other: DE99002793
  • Report No.: DOE/ER/61831--T1
  • Grant Number: FG02-94ER61831
  • DOI: 10.2172/353379 | External Link
  • Office of Scientific & Technical Information Report Number: 353379
  • Archival Resource Key: ark:/67531/metadc684234

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Office of Scientific & Technical Information Technical Reports

Reports, articles and other documents harvested from the Office of Scientific and Technical Information.

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  • December 31, 1996

Added to The UNT Digital Library

  • July 25, 2015, 2:20 a.m.

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  • April 13, 2017, 12:20 p.m.

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Ulanovsky, L. DNA sequencing technology, walking with modular primers. Final report, report, December 31, 1996; United States. (digital.library.unt.edu/ark:/67531/metadc684234/: accessed November 22, 2017), University of North Texas Libraries, Digital Library, digital.library.unt.edu; crediting UNT Libraries Government Documents Department.