Samenvatting

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.

Originele taal-2English
UitgeverijSpringer Protocols
Aantal pagina's32
ISBN van elektronische versie978-1-4939-8931-7
ISBN van geprinte versie978-1-4939-8930-0
DOI's
StatusE-pub ahead of print - 25-nov-2018

Publicatie series

NaamMethods in Molecular Biology
ISSN van geprinte versie1064-3745

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