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
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Autor*in
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
Alle
Alle
Einrichtung
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.
Erscheinungsjahr
2014
Zeitschriftentitel
Human mutation
Band
35
Ausgabe
10
Seite(n)
1260-1270
ISSN
1059-7794
Page URI
https://pub.uni-bielefeld.de/record/2691279
Zitieren
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.
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