Evaluating the Arrayed Primer Extension Resequencing Assay of TP53 Tumor Suppressor Gene
Identification of mutations in the tumor suppressor gene TP53 has implications for the molecular epidemiology and for the molecular pathology of human cancer. We have developed and evaluated an arrayed primer extension assay for covering both strands of a region of the coding sequence containing mor...
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| Published in | Proceedings of the National Academy of Sciences - PNAS Vol. 99; no. 8; pp. 5503 - 5508 |
|---|---|
| Main Authors | , , , , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
United States
National Academy of Sciences
16.04.2002
National Acad Sciences |
| Subjects | |
| Online Access | Get full text |
| ISSN | 0027-8424 1091-6490 1091-6490 |
| DOI | 10.1073/pnas.082100599 |
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| Abstract | Identification of mutations in the tumor suppressor gene TP53 has implications for the molecular epidemiology and for the molecular pathology of human cancer. We have developed and evaluated an arrayed primer extension assay for covering both strands of a region of the coding sequence containing more than 95% of the mutations described so far in TP53. On average, 97.5% of the arrayed TP53 gene sequence can be analyzed from either sense or antisense strands, and 81% from both strands. A patient DNA sample is amplified and annealed to arrayed primers, which then promote DNA polymerase extension reactions with four fluorescently labeled dideoxynucleotides. The TP53 gene chip spans exons 2-9 plus two introns from both strands. The performance of the assay was evaluated by using freshly extracted genomic DNA, as well as DNA extracted from archival (paraffin-embedded) DNA samples. The arrayed primer extension-based TP53 gene test provides an accurate and efficient tool for DNA sequence analysis of this frequently mutated gene for both research and clinical applications. |
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| AbstractList | Identification of mutations in the tumor suppressor gene TP53 has implications for the molecular epidemiology and for the molecular pathology of human cancer. We have developed and evaluated an arrayed primer extension assay for covering both strands of a region of the coding sequence containing more than 95% of the mutations described so far in TP53. On average, 97.5% of the arrayed TP53 gene sequence can be analyzed from either sense or antisense strands, and 81% from both strands. A patient DNA sample is amplified and annealed to arrayed primers, which then promote DNA polymerase extension reactions with four fluorescently labeled dideoxynucleotides. The TP53 gene chip spans exons 2–9 plus two introns from both strands. The performance of the assay was evaluated by using freshly extracted genomic DNA, as well as DNA extracted from archival (paraffin-embedded) DNA samples. The arrayed primer extension-based TP53 gene test provides an accurate and efficient tool for DNA sequence analysis of this frequently mutated gene for both research and clinical applications. APEX‖oligonucleotide array‖chip Identification of mutations in the tumor suppressor gene TP53 has implications for the molecular epidemiology and for the molecular pathology of human cancer. We have developed and evaluated an arrayed primer extension assay for covering both strands of a region of the coding sequence containing more than 95% of the mutations described so far in TP53. On average, 97.5% of the arrayed TP53 gene sequence can be analyzed from either sense or antisense strands, and 81% from both strands. A patient DNA sample is amplified and annealed to arrayed primers, which then promote DNA polymerase extension reactions with four fluorescently labeled dideoxynucleotides. The TP53 gene chip spans exons 2–9 plus two introns from both strands. The performance of the assay was evaluated by using freshly extracted genomic DNA, as well as DNA extracted from archival (paraffin-embedded) DNA samples. The arrayed primer extension-based TP53 gene test provides an accurate and efficient tool for DNA sequence analysis of this frequently mutated gene for both research and clinical applications. Identification of mutations in the tumor suppressor gene TP53 has implications for the molecular epidemiology and for the molecular pathology of human cancer. We have developed and evaluated an arrayed primer extension assay for covering both strands of a region of the coding sequence containing more than 95% of the mutations described so far in TP53. On average, 97.5% of the arrayed TP53 gene sequence can be analyzed from either sense or antisense strands 81% from both strands. A patient DNA sample is amplified and annealed to arrayed primers, which then promote DNA polymerase extension reactions with four fluorescently labeled dideoxynucleotides. The TP53 gene chip spans exons 2-9 plus two introns from both strands. The performance of the assay was evaluated by using freshly extracted genomic DNA, as well as DNA extracted from archival (paraffin-embedded) DNA samples. The arrayed primer extension-based TP53 gene test provides an accurate and efficient tool for DNA sequence analysis of this frequently mutated gene for both research and clinical applications. Identification of mutations in the tumor suppressor gene TP53 has implications for the molecular epidemiology and for the molecular pathology of human cancer. We have developed and evaluated an arrayed primer extension assay for covering both strands of a region of the coding sequence containing more than 95% of the mutations described so far in TP53. On average, 97.5% of the arrayed TP53 gene sequence can be analyzed from either sense or antisense strands, and 81% from both strands. A patient DNA sample is amplified and annealed to arrayed primers, which then promote DNA polymerase extension reactions with four fluorescently labeled dideoxynucleotides. The TP53 gene chip spans exons 2-9 plus two introns from both strands. The performance of the assay was evaluated by using freshly extracted genomic DNA, as well as DNA extracted from archival (paraffin-embedded) DNA samples. The arrayed primer extension-based TP53 gene test provides an accurate and efficient tool for DNA sequence analysis of this frequently mutated gene for both research and clinical applications. Identification of mutations in the tumor suppressor gene TP53 has implications for the molecular epidemiology and for the molecular pathology of human cancer. We have developed and evaluated an arrayed primer extension assay for covering both strands of a region of the coding sequence containing more than 95% of the mutations described so far in TP53. On average, 97.5% of the arrayed TP53 gene sequence can be analyzed from either sense or antisense strands, and 81% from both strands. A patient DNA sample is amplified and annealed to arrayed primers, which then promote DNA polymerase extension reactions with four fluorescently labeled dideoxynucleotides. The TP53 gene chip spans exons 2-9 plus two introns from both strands. The performance of the assay was evaluated by using freshly extracted genomic DNA, as well as DNA extracted from archival (paraffin-embedded) DNA samples. The arrayed primer extension-based TP53 gene test provides an accurate and efficient tool for DNA sequence analysis of this frequently mutated gene for both research and clinical applications.Identification of mutations in the tumor suppressor gene TP53 has implications for the molecular epidemiology and for the molecular pathology of human cancer. We have developed and evaluated an arrayed primer extension assay for covering both strands of a region of the coding sequence containing more than 95% of the mutations described so far in TP53. On average, 97.5% of the arrayed TP53 gene sequence can be analyzed from either sense or antisense strands, and 81% from both strands. A patient DNA sample is amplified and annealed to arrayed primers, which then promote DNA polymerase extension reactions with four fluorescently labeled dideoxynucleotides. The TP53 gene chip spans exons 2-9 plus two introns from both strands. The performance of the assay was evaluated by using freshly extracted genomic DNA, as well as DNA extracted from archival (paraffin-embedded) DNA samples. The arrayed primer extension-based TP53 gene test provides an accurate and efficient tool for DNA sequence analysis of this frequently mutated gene for both research and clinical applications. Identification of mutations in the tumor suppressor gene TP53 has implications for the molecular epidemiology and for the molecular pathology of human cancer. Tonisson et al developed and evaluated an arrayed primer extension assay for covering both strands of a region of the coding sequence containing more than 95% of the mutations described so far in TP53. |
| Author | Meiel, Aune Zernant, Jana Kurg, Ants Slavin, Georg Roomere, Hanno Pavel, Hendrik Hainaut, Pierre Tõnisson, Neeme Metspalu, Andres |
| AuthorAffiliation | Asper, Ltd., 3 Oru Street, 51014 Tartu, Estonia; † Institute of Molecular and Cell Biology, University of Tartu/Estonian Biocentre, 23 Riia Street, 51010 Tartu, Estonia; and ‡ International Agency for Research on Cancer, 150, Cours Albert Thomas, F-69372 Lyon Cedex 08, France |
| AuthorAffiliation_xml | – name: Asper, Ltd., 3 Oru Street, 51014 Tartu, Estonia; † Institute of Molecular and Cell Biology, University of Tartu/Estonian Biocentre, 23 Riia Street, 51010 Tartu, Estonia; and ‡ International Agency for Research on Cancer, 150, Cours Albert Thomas, F-69372 Lyon Cedex 08, France |
| Author_xml | – sequence: 1 givenname: Neeme surname: Tõnisson fullname: Tõnisson, Neeme – sequence: 2 givenname: Jana surname: Zernant fullname: Zernant, Jana – sequence: 3 givenname: Ants surname: Kurg fullname: Kurg, Ants – sequence: 4 givenname: Hendrik surname: Pavel fullname: Pavel, Hendrik – sequence: 5 givenname: Georg surname: Slavin fullname: Slavin, Georg – sequence: 6 givenname: Hanno surname: Roomere fullname: Roomere, Hanno – sequence: 7 givenname: Aune surname: Meiel fullname: Meiel, Aune – sequence: 8 givenname: Pierre surname: Hainaut fullname: Hainaut, Pierre – sequence: 9 givenname: Andres surname: Metspalu fullname: Metspalu, Andres |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/11960007$$D View this record in MEDLINE/PubMed |
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| Snippet | Identification of mutations in the tumor suppressor gene TP53 has implications for the molecular epidemiology and for the molecular pathology of human cancer.... Identification of mutations in the tumor suppressor gene TP53 has implications for the molecular epidemiology and for the molecular pathology of human cancer.... |
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| SubjectTerms | 53 gene Algorithms Biological Sciences Cancer Codon Codons DNA DNA Mutational Analysis - methods Epidemiology Exons Genes Genes, p53 - genetics Genetic mutation Genetic Techniques Humans Introns Missense mutation Mutation Mutation, Missense Oligonucleotide Array Sequence Analysis Oligonucleotides p53 genes Sequencing TP53 gene Tumors |
| Title | Evaluating the Arrayed Primer Extension Resequencing Assay of TP53 Tumor Suppressor Gene |
| URI | https://www.jstor.org/stable/3058522 http://www.pnas.org/content/99/8/5503.abstract https://www.ncbi.nlm.nih.gov/pubmed/11960007 https://www.proquest.com/docview/201394124 https://www.proquest.com/docview/18314978 https://www.proquest.com/docview/71612458 https://pubmed.ncbi.nlm.nih.gov/PMC122799 http://doi.org/10.1073/pnas.082100599 |
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