Single G2/M Nucleus Sequencing of Cells in S Phase/Single Nucleus Exome Sequencing

A modified MDA protocol, nuc-seq takes advantage of the fact that a single cell in the G2_M stage of the cell cycle has 4 copies of the genome. This property allows the cells to be isolated with a cell sorter, and it also significantly increases the genome coverage of single cells (Wang et al., 2014). SNES is an additional variation that includes targeted selection and sequencing of the exomes (Leung et al., 2015). Div-Seq is a variation that combines nuc-seq with pulse labeling of proliferating cells by 5-ethynyl-2_-deoxyuridine (EdU) Habib et al., 2016)


  • nuc-seq: Improves physical coverage performance to more than 90% for single-cell sequencing
  • SNES: 95.94% exome coverage in single cells
  • SNES: Detection efficiencies of 92.37% for SNVs in an isogenic population


  • nuc-seq: Cannot be applied to cells with low proliferation rates
  • SNES: Limited to exomes


Illumina Library prep and Array Kit Selector


Zhang X., Marjani S. L., Hu Z., Weissman S. M., Pan X., et al. Single-Cell Sequencing for Precise Cancer Research: Progress and Prospects. Cancer Res. 2016;76:1305-1312

Leung M. L., Wang Y., Waters J. and Navin N. E. SNES: single nucleus exome sequencing. Genome Biol. 2015;16:55

Saadatpour A., Lai S., Guo G. and Yuan G. C. Single-Cell Analysis in Cancer Genomics. Trends Genet. 2015;31:576-586

Szulwach K. E., Chen P., Wang X., et al. Single-Cell Genetic Analysis Using Automated Microfluidics to Resolve Somatic Mosaicism. PLoS One. 2015;10:e0135007

Voet T. and Van Loo P. SNES makes sense? Single-cell exome sequencing evolves. Genome Biol. 2015;16:86

Wang Y. and Navin N. E. Advances and applications of single-cell sequencing technologies. Mol Cell. 2015;58:598-609


Leung M. L., Wang Y., Waters J. and Navin N. E. SNES: single nucleus exome sequencing. Genome Biol. 2015;16:55

Leung M. L., Wang Y., Kim C., Gao R., Jiang J., et al. Highly multiplexed targeted DNA sequencing from single nuclei. Nat Protoc. 2016;11:214-235

Habib N., Li Y., Heidenreich M., et al. Div-Seq: A single nucleus RNA-Seq method reveals dynamics of rare adult newborn neurons in the CNS. bioRxiv. 2016;