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Whole Genome Amplification in Genomic Analysis of Single Circulating Tumor Cells

  • Christin Gasch
  • Klaus Pantel
  • Sabine RiethdorfEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1347)

Abstract

Investigation of the genome of organisms is one of the major basics in molecular biology to understand the complex organization of cells. While genomic DNA can easily be isolated from tissues or cell cultures of plant, animal or human origin, DNA extraction from single cells is still challenging. Here, we describe three techniques for the amplification of genomic DNA from fixed single circulating tumor cells (CTC) isolated from blood of cancer patients. This amplification is aimed to increase DNA amounts from those of one cell to yields sufficient for different DNA analyses such as mutational analysis including next-generation sequencing, array-comparative genome hybridization (CGH), and quantitative measurement of gene amplifications. Molecular analysis of CTC as liquid biopsy can be used to identify therapeutic targets in personalized medicine directed, e.g. against human epidermal growth factor receptor 2 (HER2) or epidermal growth factor receptor (EGFR) and to stratify the patients to those therapies.

Keywords

Whole genome amplification Single cell analysis Circulating tumor cells Multiple-strand displacement amplification Linker-adapter PCR Genomic analysis 

Notes

Acknowledgement

This work was supported by the European Community’s 7th Framework Programme (FP7/2007–2013) under grant agreement no. 202230, acronym GENINCA to KP, and the ERC-2010-AdG_20100317 DISSECT to KP.

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Christin Gasch
    • 1
  • Klaus Pantel
    • 1
  • Sabine Riethdorf
    • 1
    Email author
  1. 1.Department of Tumor BiologyUniversity Medical Center Hamburg-EppendorfHamburgGermany

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