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Heat-Induced Fragmentation and Adapter-Assisted Whole Genome Amplification Using GenomePlex® Single-Cell Whole Genome Amplification Kit (WGA4)

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Whole Genome Amplification

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1347))

Abstract

Whole genome amplification (WGA) is a widely used technique allowing multiplying picogram amounts of target DNA by several orders of magnitude. The technique described here is based on heat-induced random fragmentation yielding DNA strands mainly ranging from 0.1 to 1 kb in length. The fragmented DNA is then subjected to library generation by annealing of adaptor sequences to both ends of the DNA fragments. Using primers hybridizing to the adapter sequences, the DNA is amplified by thermal cycling. This amplification typically yields > 2 mg DNA from a single cell, is suited for amplifying DNA isolated from (partly) degraded samples [e.g. formalin-fixed paraffin-embedded (FFPE) material] and works well when used for array-comparative genome hybridization (array-CGH).

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Acknowledgement

This work was supported by the EU SAFE Network of Excellence (LSHB-CT-2004-503243, EU 6th Framework Package) and the County of Styria, Austria.

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Authors and Affiliations

  1. Research Unit for Single Cell Analysis, Institute of Cell Biology, Histology & Embryology, Medical University of Graz, Harrachgasse 21, Graz, 8010, Austria

    Amin El-Heliebi, Shukun Chen & Thomas Kroneis

  2. Sahlgrenska Cancer Center, University of Gothenburg, Gothenburg, Sweden

    Thomas Kroneis

Authors
  1. Amin El-Heliebi
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  2. Shukun Chen
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  3. Thomas Kroneis
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Corresponding author

Correspondence to Thomas Kroneis .

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Editors and Affiliations

  1. Institute of Cell Biology, Histology & Embryology, Medical University of Graz, Graz, Austria

    Thomas Kroneis

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El-Heliebi, A., Chen, S., Kroneis, T. (2015). Heat-Induced Fragmentation and Adapter-Assisted Whole Genome Amplification Using GenomePlex® Single-Cell Whole Genome Amplification Kit (WGA4). In: Kroneis, T. (eds) Whole Genome Amplification. Methods in Molecular Biology, vol 1347. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2990-0_7

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  • DOI: https://doi.org/10.1007/978-1-4939-2990-0_7

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2989-4

  • Online ISBN: 978-1-4939-2990-0

  • eBook Packages: Springer Protocols

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