Temperature scans/cycles for the detection of low abundant DNA point mutations on microarrays |
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Authors: | Pingel Julia Buhot Arnaud Calemczuk Roberto Livache Thierry |
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Affiliation: | INAC, SPrAM (UMR 5819, CEA, CNRS, UJF), CEA Grenoble, 17 rue des Martyrs, F-38054 Grenoble cedex 9, France. |
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Abstract: | The possibility to detect low abundant DNA point mutations is essential for early cancer diagnosis and/or prognosis. Furthermore, in order to be less invasive, the somatic mutations are not only sought in tumor extract samples but also from body fluids or stools rendering their content even more diluted compared to the wild type sequences. In this short communication, we propose two protocols based on temperature scans or cycles for the enrichment of the mutation strands hybridized on microarrays. We predict numerically and confirm experimentally a 10-fold increase in the fraction of mutated DNA hybridized on the microarray compared to the sample content. Coupled to more standard solution phase enrichment techniques, it would be possible to lower by one order of magnitude the current detection limit with the advantage of multiple mutation detections offered by the microarray technology. |
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