Method for priming and DNA sequencing

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Description

A method is presented for improving the priming specificity of an oligonucleotide primer that is non-unique in a nucleic acid template which includes selecting a continuous stretch of several nucleotides in the template DNA where one of the four bases does not occur in the stretch. This also includes bringing the template DNA in contract with a non-unique primer partially or fully complimentary to the sequence immediately upstream of the selected sequence stretch. This results in polymerase-mediated differential extension of the primer in the presence of a subset of deoxyribonucleotide triphosphates that does not contain the base complementary to the ... continued below

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Medium: P; Size: 36 p.

Creation Information

Mugasimangalam, R.C. & Ulanovsky, L.E. December 1, 1997.

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Description

A method is presented for improving the priming specificity of an oligonucleotide primer that is non-unique in a nucleic acid template which includes selecting a continuous stretch of several nucleotides in the template DNA where one of the four bases does not occur in the stretch. This also includes bringing the template DNA in contract with a non-unique primer partially or fully complimentary to the sequence immediately upstream of the selected sequence stretch. This results in polymerase-mediated differential extension of the primer in the presence of a subset of deoxyribonucleotide triphosphates that does not contain the base complementary to the base absent in the selected sequence stretch. These reactions occur at a temperature sufficiently low for allowing the extension of the non-unique primer. The method causes polymerase-mediated extension reactions in the presence of all four natural deoxyribonucleotide triphosphates or modifications. At this high temperature discrimination occurs against priming sites of the non-unique primer where the differential extension has not made the primer sufficiently stable to prime. However, the primer extended at the selected stretch is sufficiently stable to prime.

Physical Description

Medium: P; Size: 36 p.

Notes

OSTI as DE98001644

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  • Other Information: PBD: 1997

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  • Other: DE98001644
  • Report No.: PATENTS-US--A-PCT/US97/17305
  • Grant Number: FG02-94ER61831
  • Office of Scientific & Technical Information Report Number: 555434
  • Archival Resource Key: ark:/67531/metadc691210

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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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Creation Date

  • December 1, 1997

Added to The UNT Digital Library

  • Aug. 14, 2015, 8:43 a.m.

Description Last Updated

  • Nov. 6, 2015, 4:01 p.m.

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Mugasimangalam, R.C. & Ulanovsky, L.E. Method for priming and DNA sequencing, patent, December 1, 1997; United States. (digital.library.unt.edu/ark:/67531/metadc691210/: accessed October 15, 2018), University of North Texas Libraries, Digital Library, digital.library.unt.edu; crediting UNT Libraries Government Documents Department.