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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
1
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pubmed:dateCreated |
1997-3-24
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pubmed:abstractText |
Double-stranded DNA fragments of different length with and without the incorporation of 7-deazapurine deoxynucleotides have been prepared via the polymerase chain reaction (PCR) using exo(-)Pseudococcus furiosus DNA polymerase, unmodified primers, and c7-dATP and c7-dGTP as the only purine triphosphates. In spite of the presence of some unmodified purine moieties due to the primers, matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry revealed remarkable differences between the modified and unmodified DNA fragments. The incorporation of 7-deazapurine nucleotides resulted in a significant reduction of fragmentation and therefore in increased signal intensities and higher mass resolution. The molecular weights could be determined with an accuracy of up to 0.03%. Mass resolution was sufficient to resolve the (M + H)+ signals of the two single strands of a 7-deazapurine-containing 99-base-pair DNA duplex. Thus, MALDI-TOF mass spectrometry offers a very fast and accurate way to detect and analyze short PCR products sufficient in length for many diagnostic applications without gel electrophoresis and labeling.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Dec
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pubmed:issn |
0003-2697
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
1
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pubmed:volume |
243
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
55-65
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pubmed:dateRevised |
2000-12-18
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pubmed:meshHeading |
pubmed-meshheading:8954525-Autoradiography,
pubmed-meshheading:8954525-Base Sequence,
pubmed-meshheading:8954525-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:8954525-Molecular Sequence Data,
pubmed-meshheading:8954525-Polymerase Chain Reaction,
pubmed-meshheading:8954525-Purines,
pubmed-meshheading:8954525-Sequence Analysis, DNA,
pubmed-meshheading:8954525-Spectrometry, Mass, Matrix-Assisted Laser...
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pubmed:year |
1996
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pubmed:articleTitle |
Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for the detection of polymerase chain reaction products containing 7-deazapurine moieties.
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pubmed:affiliation |
Department of Biochemistry and Molecular Biology, Faculty of Chemistry, University of Hamburg, Germany.
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pubmed:publicationType |
Journal Article
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