Abstract
High-throughput next generation sequencing karyotyping has emerged as a powerful tool for the detection of genomic heterogeneity in normal tissues and cancers. Here we describe a single-cell whole genome sequencing (scWGS) platform to assess whole-chromosome aneuploidy, structural aneuploidies involving only chromosome fragments and more local small copy number alterations in individual cells. We provide a detailed protocol for the isolation, library preparation, low coverage sequencing and data analysis of single cells. Since our approach does not involve a whole-genome preamplification step, our method allows for acquisition of reliable high-resolution single-cell copy number profiles. Moreover, the protocol allows multiplexing of 384 single-cell libraries in one sequencing run, thereby significantly reducing sequencing costs and can be completed in 3–4 days starting from single cell isolation to analysis of sequencing data.
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References
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Acknowledgments
This work was supported by a Dutch Cancer Society grant (KWF grant RUG-2012-5549) and a Groningen Foundation for Paediatric Oncology (SKOG) grant to FF and PML, an Advanced ERC grant to PML, and by an NWO (The Netherlands Organisation for Scientific Research) MEERVOUD grant and a University of Groningen Rosalind Franklin grant to MCT.
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van den Bos, H. et al. (2019). Quantification of Aneuploidy in Mammalian Systems. In: Demaria, M. (eds) Cellular Senescence. Methods in Molecular Biology, vol 1896. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8931-7_15
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DOI: https://doi.org/10.1007/978-1-4939-8931-7_15
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Publisher Name: Humana Press, New York, NY
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