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. 2018 Jul 9;46(19):e112. doi: 10.1093/nar/gky590

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© The Author(s) 2018. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 6. — Application of ddPCR to determine relative DNA replication time in human cells. (A) DNA copy number analysis in non-replicating human cells. DNA from male MRC-5 cells arrested in G1 phase of the cell cycle was analysed by ddPCR with probes targeting sex chromosomes, autosomal chromosomes and duplicated genes. Relative copy number was calculated as a mean concentration of all probes adjusted by copy number. (B) DNA concentration analysed by ddPCR across a known replication timing transition zone on chromosome 18. Top: non-replicating control from FACS-enriched G1 HeLa cells, as well as FACS-enriched S phase HeLa cells analysed by ddPCR and sortSeq. Note that G1 and S phase samples, while coming from the same initial population of cells, are independent DNA samples and their absolute concentration cannot be compared. Bottom: repli-seq data of the corresponding genomic region from (14,41). cPNS - combined percentage-normalised signal.