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Automated array-CGH optimized for archival formalin-fixed, paraffin-embedded tumor material

BACKGROUND: Array Comparative Genomic Hybridization (aCGH) is a rapidly evolving technology that still lacks complete standardization. Yet, it is of great importance to obtain robust and reproducible data to enable meaningful multiple hybridization comparisons. Special difficulties arise when aCGH i...

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Detalles Bibliográficos
Autores principales: Joosse, Simon A, van Beers, Erik H, Nederlof, Petra M
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2007
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1829401/
https://www.ncbi.nlm.nih.gov/pubmed/17343727
http://dx.doi.org/10.1186/1471-2407-7-43
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author Joosse, Simon A
van Beers, Erik H
Nederlof, Petra M
author_facet Joosse, Simon A
van Beers, Erik H
Nederlof, Petra M
author_sort Joosse, Simon A
collection PubMed
description BACKGROUND: Array Comparative Genomic Hybridization (aCGH) is a rapidly evolving technology that still lacks complete standardization. Yet, it is of great importance to obtain robust and reproducible data to enable meaningful multiple hybridization comparisons. Special difficulties arise when aCGH is performed on archival formalin-fixed, paraffin-embedded (FFPE) tissue due to its variable DNA quality. Recently, we have developed an effective DNA quality test that predicts suitability of archival samples for BAC aCGH. METHODS: In this report, we first used DNA from a cancer cell-line (SKBR3) to optimize the aCGH protocol for automated hybridization, and subsequently optimized and validated the procedure for FFPE breast cancer samples. We aimed for highest throughput, accuracy, and reproducibility applicable to FFPE samples, which can also be important in future diagnostic use. RESULTS: Our protocol of automated array-CGH on archival FFPE ULS-labeled DNA showed very similar results compared with published data and our previous manual hybridization method. CONCLUSION: This report combines automated aCGH on unamplified archival FFPE DNA using non-enzymatic ULS labeling, and describes an optimized protocol for this combination resulting in improved quality and reproducibility.
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spelling pubmed-18294012007-03-22 Automated array-CGH optimized for archival formalin-fixed, paraffin-embedded tumor material Joosse, Simon A van Beers, Erik H Nederlof, Petra M BMC Cancer Technical Advance BACKGROUND: Array Comparative Genomic Hybridization (aCGH) is a rapidly evolving technology that still lacks complete standardization. Yet, it is of great importance to obtain robust and reproducible data to enable meaningful multiple hybridization comparisons. Special difficulties arise when aCGH is performed on archival formalin-fixed, paraffin-embedded (FFPE) tissue due to its variable DNA quality. Recently, we have developed an effective DNA quality test that predicts suitability of archival samples for BAC aCGH. METHODS: In this report, we first used DNA from a cancer cell-line (SKBR3) to optimize the aCGH protocol for automated hybridization, and subsequently optimized and validated the procedure for FFPE breast cancer samples. We aimed for highest throughput, accuracy, and reproducibility applicable to FFPE samples, which can also be important in future diagnostic use. RESULTS: Our protocol of automated array-CGH on archival FFPE ULS-labeled DNA showed very similar results compared with published data and our previous manual hybridization method. CONCLUSION: This report combines automated aCGH on unamplified archival FFPE DNA using non-enzymatic ULS labeling, and describes an optimized protocol for this combination resulting in improved quality and reproducibility. BioMed Central 2007-03-07 /pmc/articles/PMC1829401/ /pubmed/17343727 http://dx.doi.org/10.1186/1471-2407-7-43 Text en Copyright © 2007 Joosse et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( (http://creativecommons.org/licenses/by/2.0) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Technical Advance
Joosse, Simon A
van Beers, Erik H
Nederlof, Petra M
Automated array-CGH optimized for archival formalin-fixed, paraffin-embedded tumor material
title Automated array-CGH optimized for archival formalin-fixed, paraffin-embedded tumor material
title_full Automated array-CGH optimized for archival formalin-fixed, paraffin-embedded tumor material
title_fullStr Automated array-CGH optimized for archival formalin-fixed, paraffin-embedded tumor material
title_full_unstemmed Automated array-CGH optimized for archival formalin-fixed, paraffin-embedded tumor material
title_short Automated array-CGH optimized for archival formalin-fixed, paraffin-embedded tumor material
title_sort automated array-cgh optimized for archival formalin-fixed, paraffin-embedded tumor material
topic Technical Advance
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1829401/
https://www.ncbi.nlm.nih.gov/pubmed/17343727
http://dx.doi.org/10.1186/1471-2407-7-43
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