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  Genome-wide massively parallel sequencing of formaldehyde fixed-paraffin embedded (FFPE) tumor tissues for copy-number- and mutation-analysis.

Schweiger, M. R., Kerick, M., Timmermann, B., Albrecht, M. W., Borodina, T., Parkhomchuk, D., et al. (2009). Genome-wide massively parallel sequencing of formaldehyde fixed-paraffin embedded (FFPE) tumor tissues for copy-number- and mutation-analysis. PLoS ONE, 4(5), e5548-e5548. doi:10.1371/journal.pone.0005548.

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Genre: Journal Article
Alternative Title : PLOS ONE

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journal.pone.0005548.pdf (Any fulltext), 576KB
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 Creators:
Schweiger, Michal R.1, Author           
Kerick, Martin1, Author           
Timmermann, Bernd2, Author           
Albrecht, Marcus W.3, Author
Borodina, Tatjana4, Author           
Parkhomchuk, Dmitri5, Author           
Zatloukal, Kurt, Author
Lehrach, Hans5, Author           
Affiliations:
1Cancer Genomics (Michal-Ruth Schweiger), Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1479649              
2Sequencing, Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1433559              
3Max Planck Society, ou_persistent13              
4Technology Development(Alexey Soldatov), Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1479657              
5Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1433550              

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 Abstract: Cancer re-sequencing programs rely on DNA isolated from fresh snap frozen tissues, the preparation of which is combined with additional preservation efforts. Tissue samples at pathology departments are routinely stored as formalin-fixed and paraffin-embedded (FFPE) samples and their use would open up access to a variety of clinical trials. However, FFPE preparation is incompatible with many down-stream molecular biology techniques such as PCR based amplification methods and gene expression studies. Methodology/Principal Findings Here we investigated the sample quality requirements of FFPE tissues for massively parallel short-read sequencing approaches. We evaluated key variables of pre-fixation, fixation related and post-fixation processes that occur in routine medical service (e.g. degree of autolysis, duration of fixation and of storage). We also investigated the influence of tissue storage time on sequencing quality by using material that was up to 18 years old. Finally, we analyzed normal and tumor breast tissues using the Sequencing by Synthesis technique (Illumina Genome Analyzer, Solexa) to simultaneously localize genome-wide copy number alterations and to detect genomic variations such as substitutions and point-deletions and/or insertions in FFPE tissue samples. Conclusions/Significance The application of second generation sequencing techniques on small amounts of FFPE material opens up the possibility to analyze tissue samples which have been collected during routine clinical work as well as in the context of clinical trials. This is in particular important since FFPE samples are amply available from surgical tumor resections and histopathological diagnosis, and comprise tissue from precursor lesions, primary tumors, lymphogenic and/or hematogenic metastases. Large-scale studies using this tissue material will result in a better prediction of the prognosis of cancer patients and the early identification of patients which will respond to therapy.

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Language(s): eng - English
 Dates: 2009-05-14
 Publication Status: Published in print
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Title: PLoS ONE
  Alternative Title : PLOS ONE
Source Genre: Journal
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Pages: - Volume / Issue: 4 (5) Sequence Number: - Start / End Page: e5548 - e5548 Identifier: ISSN: 1932-6203