A New Workflow for Whole-Genome Sequencing of Single Human Cells

Binder V, Bartenhagen C, Okpanyi V, Gombert M, Moehlendick B, Behrens B, Klein H-U, Rieder H, Krell P, Dugas M, Stoecklein NH, et al. (2014)
Human mutation 35(10): 1260-1270.

Zeitschriftenaufsatz | Veröffentlicht | Englisch
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Binder, Vera; Bartenhagen, Christoph; Okpanyi, Vera; Gombert, Michael; Moehlendick, Birte; Behrens, Bianca; Klein, Hans-Ulrich; Rieder, Harald; Krell, PinaUniBi; Dugas, Martin; Stoecklein, Nikolas Hendrik; Borkhardt, Arndt
Abstract / Bemerkung
: Unbiased amplification of the whole-genome amplification (WGA) of single cells is crucial to study cancer evolution and genetic heterogeneity, but is challenging due to the high complexity of the human genome. Here, we present a new workflow combining an efficient adapter-linker PCR-based WGA method with second-generation sequencing. This approach allows comparison of single cells at base pair resolution. Amplification recovered up to 74% of the human genome. Copy-number variants and loss of heterozygosity detected in single cell genomes showed concordance of up to 99% to pooled genomic DNA. Allele frequencies of mutations could be determined accurately due to an allele dropout rate of only 2%, clearly demonstrating the low bias of our PCR-based WGA approach. Sequencing with paired-end reads allowed genome-wide analysis of structural variants. By direct comparison to other WGA methods, we further endorse its suitability to analyze genetic heterogeneity.
Human mutation
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Binder V, Bartenhagen C, Okpanyi V, et al. A New Workflow for Whole-Genome Sequencing of Single Human Cells. Human mutation. 2014;35(10):1260-1270.
Binder, V., Bartenhagen, C., Okpanyi, V., Gombert, M., Moehlendick, B., Behrens, B., Klein, H. - U., et al. (2014). A New Workflow for Whole-Genome Sequencing of Single Human Cells. Human mutation, 35(10), 1260-1270. doi:10.1002/humu.22625
Binder, Vera, Bartenhagen, Christoph, Okpanyi, Vera, Gombert, Michael, Moehlendick, Birte, Behrens, Bianca, Klein, Hans-Ulrich, et al. 2014. “A New Workflow for Whole-Genome Sequencing of Single Human Cells”. Human mutation 35 (10): 1260-1270.
Binder, V., Bartenhagen, C., Okpanyi, V., Gombert, M., Moehlendick, B., Behrens, B., Klein, H. - U., Rieder, H., Krell, P., Dugas, M., et al. (2014). A New Workflow for Whole-Genome Sequencing of Single Human Cells. Human mutation 35, 1260-1270.
Binder, V., et al., 2014. A New Workflow for Whole-Genome Sequencing of Single Human Cells. Human mutation, 35(10), p 1260-1270.
V. Binder, et al., “A New Workflow for Whole-Genome Sequencing of Single Human Cells”, Human mutation, vol. 35, 2014, pp. 1260-1270.
Binder, V., Bartenhagen, C., Okpanyi, V., Gombert, M., Moehlendick, B., Behrens, B., Klein, H.-U., Rieder, H., Krell, P., Dugas, M., Stoecklein, N.H., Borkhardt, A.: A New Workflow for Whole-Genome Sequencing of Single Human Cells. Human mutation. 35, 1260-1270 (2014).
Binder, Vera, Bartenhagen, Christoph, Okpanyi, Vera, Gombert, Michael, Moehlendick, Birte, Behrens, Bianca, Klein, Hans-Ulrich, Rieder, Harald, Krell, Pina, Dugas, Martin, Stoecklein, Nikolas Hendrik, and Borkhardt, Arndt. “A New Workflow for Whole-Genome Sequencing of Single Human Cells”. Human mutation 35.10 (2014): 1260-1270.

9 Zitationen in Europe PMC

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