[One-step PCR sequencing]. Final report, July 1, 1994--August 31, 1997

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The author explored the use of a novel class of boronated nucleic acids, the boranophosphates, as an alternative, but complementary method to dideoxysequencing. Boranophosphates can be used to directly amplify and sequence single- or double-stranded DNA. Fragments are derived not from truncations during polymerase synthesis, but from insertion and digestion back to a boronated marker or delimiter that was incorporated during exponential amplification. The method, which the author calls Boronated One-Step PCR Sequencing, is unique in that it employs a new class of {alpha}-P-boronated 2{prime}-deoxynucleoside 5{prime}-triphosphates first synthesized in the laboratory. These boronated triphosphates exhibit useful properties: (a) they are ... continued below

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8 p.

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Shaw, B.R. December 31, 1997.

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Description

The author explored the use of a novel class of boronated nucleic acids, the boranophosphates, as an alternative, but complementary method to dideoxysequencing. Boranophosphates can be used to directly amplify and sequence single- or double-stranded DNA. Fragments are derived not from truncations during polymerase synthesis, but from insertion and digestion back to a boronated marker or delimiter that was incorporated during exponential amplification. The method, which the author calls Boronated One-Step PCR Sequencing, is unique in that it employs a new class of {alpha}-P-boronated 2{prime}-deoxynucleoside 5{prime}-triphosphates first synthesized in the laboratory. These boronated triphosphates exhibit useful properties: (a) they are heat stable, (b) they can be base-specifically incorporated into DNA during the polymerase chain reaction, and (c) once incorporated, the boranophosphate nucleotide (marker) blocks the action of exonuclease. Thus, the positions of the stably-incorporated boronated markers can be revealed by a simple exonuclease digestion, producing a series of fragments--each of which is terminated base-specifically at the boronated markers--and thereby defining the sequence of the PCR product.

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8 p.

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OSTI as DE99002794

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  • Other Information: PBD: [1997]

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  • Other: DE99002794
  • Report No.: DOE/ER/61882--T1
  • Grant Number: FG05-94ER61882
  • DOI: 10.2172/353382 | External Link
  • Office of Scientific & Technical Information Report Number: 353382
  • Archival Resource Key: ark:/67531/metadc686260

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  • December 31, 1997

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  • July 25, 2015, 2:20 a.m.

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  • Nov. 6, 2015, 12:56 p.m.

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Shaw, B.R. [One-step PCR sequencing]. Final report, July 1, 1994--August 31, 1997, report, December 31, 1997; United States. (digital.library.unt.edu/ark:/67531/metadc686260/: accessed September 23, 2017), University of North Texas Libraries, Digital Library, digital.library.unt.edu; crediting UNT Libraries Government Documents Department.