Protocol

Genomic Structural Variants

Volume 838 of the series Methods in Molecular Biology pp 329-341

Date:

High-Resolution Copy Number Profiling by Array CGH Using DNA Isolated from Formalin-Fixed, Paraffin-Embedded Tissues

  • Hendrik F. van EssenAffiliated withUniversity of Antwerp (UA)
  • , Bauke YlstraAffiliated withVU Medical Center Email author 

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Abstract

We describe protocols to acquire high-quality DNA from formalin-fixed, paraffin-embedded (FFPE) tissues for the use in array comparative genome hybridization (CGH). Formalin fixation combined with paraffin embedding is routine procedure for solid malignancies in the diagnostic practice of the pathologist. As a consequence, large archives of FFPE tissues are available in pathology institutes across the globe. This archival material is for many research questions an invaluable resource, with long-term clinical follow-up and survival data available. FFPE is, thus, highly attractive for large genomics studies, including experiments requiring samples for test/learning and validation. Most larger array CGH studies have, therefore, made use of FFPE material and show that CNAs have tumor- and tissue-specific traits (Chin et al. Cancer Cell 10: 529–541, 2006; Fridlyand et al. BMC Cancer 6: 96, 2006; Weiss et al. Oncogene 22: 1872–1879, 2003; Jong et al. Oncogene 26: 1499–1506, 2007). The protocols described are tailored to array CGH of FFPE solid malignancies: from sectioning FFPE blocks to specific cynosures for pathological revisions of sections, DNA isolation, quality testing, and amplification. The protocols are technical in character and elaborate up to the labeling of isolated DNA while further processes and interpretation and data analysis are beyond the scope.

Key words

Formalin fixed, paraffin embedded Chromosomal DNA Microarray Copy number aberrations Archival tissue Sodium thiocyanate Array comparative genome hybridization