A Novel Multiplexing, Polymerase Chain Reaction-Based Assay for the Analysis of Chromosome 18q Status in Colorectal Cancer Nadina Erill, Anna Colomer,

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A Novel Multiplexing, Polymerase Chain Reaction-Based Assay for the Analysis of Chromosome 18q Status in Colorectal Cancer Nadina Erill, Anna Colomer, Miquel Calvo, August Vidal, Ruth Román, Montse Verdú, Carlos Cordón-Cardó, Xavier Puig The Journal of Molecular Diagnostics Volume 7, Issue 4, Pages 478-485 (October 2005) DOI: 10.1016/S1525-1578(10)60578-8 Copyright © 2005 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 1 Representative electropherograms for the five multiplexed chromosome 18q markers. The fluorescent labels were chosen such that the amplification products were sufficiently separated in size to prevent overlapping, and primer molar concentrations were also adjusted to achieve similar peak amplitudes for all markers. The Journal of Molecular Diagnostics 2005 7, 478-485DOI: (10.1016/S1525-1578(10)60578-8) Copyright © 2005 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 2 GeneScan electropherograms of two different microsatellites using three decreasing DNA concentrations. For each informative marker, the major peak of each allele selected for LOH analysis, consistent with highest one, is pointed by an arrow. The relative ratios between normal and tumor allele for each experiment are exposed. The Journal of Molecular Diagnostics 2005 7, 478-485DOI: (10.1016/S1525-1578(10)60578-8) Copyright © 2005 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 3 Polyacrylamide gel images and their corresponding electropherograms of marker D18S55. Representative cases for three different results are shown. The results given by the two methods in examples A and B are coincident, whereas the ambiguous result of acrylamide gel shown in example C is resolved by GeneScan analysis. The Journal of Molecular Diagnostics 2005 7, 478-485DOI: (10.1016/S1525-1578(10)60578-8) Copyright © 2005 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 4 Microsatellite markers used for LOH determination at the long arm of chromosome 18 in stage II colorectal tumors. Revision of relevant results previously described. The frequencies of allelic loss detected with each panel of markers, altogether with the reference of the study are indicated. *An additional intragenic microsatellite located within the DCC (18qDCC-TA) is evaluated. **Inferior lane corresponds to the present study. The Journal of Molecular Diagnostics 2005 7, 478-485DOI: (10.1016/S1525-1578(10)60578-8) Copyright © 2005 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions