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Deterministic Whole-Genome Amplification of Single Cells

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

Abstract

This chapter describes a single-cell whole genome amplification method (WGA) that has been originally published under the name “Single Cell Comparative Genomic Hybridization (SCOMP)” (Klein et al., Proc Natl Acad Sci U S A 96(8):4494–4499, 1999). The method has recently become available commercially under the name “Ampli1 WGA Kit.” It is a PCR-based technique for whole genome amplification (WGA) allowing comprehensive and quite uniform amplification of DNA from low quantities of input DNA material, in particular single cells. The method is based on a ligation-mediated adaptor linker PCR approach. In contrast to other PCR-based WGA approaches, both the primer design and mechanism underlying the fragmentation of genome are nonrandom, enabling high priming efficiency and deterministic fragmentation of template DNA. This is particularly important for the design of (diagnostic) assays targeting specific loci. Here, we describe the WGA protocol for amplification of single-cell genomes designed to provide high-quality material in quantity sufficient for a number of locus-specific and genome-wide downstream assays [e.g., targeted Sanger sequencing, restriction fragment length polymorphism (RFLP), quantitative PCR (qPCR), and array comparative genomic hybridization (CGH)].

Key words

  • Whole genome amplification (WGA)
  • Single-cell analysis
  • Multiple displacement amplification (MDA)
  • Ligation-mediated PCR

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  • DOI: 10.1007/978-1-4939-2990-0_5
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Correspondence to Christoph A. Klein .

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Czyż, Z.T., Klein, C.A. (2015). Deterministic Whole-Genome Amplification of Single Cells. 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_5

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  • DOI: https://doi.org/10.1007/978-1-4939-2990-0_5

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2989-4

  • Online ISBN: 978-1-4939-2990-0

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