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Quantitative analysis of gene expression in a single cell by qPCR

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Abstract

We developed a quantitative PCR method featuring a reusable single-cell cDNA library immobilized on beads for measuring the expression of multiple genes in a single cell. We used this method to analyze multiple cDNA targets (from several copies to several hundred thousand copies) with an experimental error of 15.9% or less. This method is sufficiently accurate to investigate the heterogeneity of single cells.

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Figure 1: Efficiency of cDNA production.
Figure 2: Characteristics of qPCR with beads.
Figure 3: Number of cDNA molecules measured in single cells and cell pools.
Figure 4: Change in standard deviation of the number of target cDNA molecules.

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References

  1. Levsky, J.M. & Singer, R.H. Trends Cell Biol. 13, 4–6 (2003).

    Article  CAS  Google Scholar 

  2. Kamme, F. et al. J. Neurosci. 23, 3607–3615 (2003).

    Article  CAS  Google Scholar 

  3. Kurimoto, K. et al. Nucleic Acids Res. 34, e42 (2006).

    Article  Google Scholar 

  4. Ginsberg, S.D., Che, S., Counts, S.E. & Mufson, E.J. NeuroRx 3, 302–318 (2006).

    Article  CAS  Google Scholar 

  5. Hartshorn, C., Anshelevich, A. & Wangh, L.J. BMC Biotechnol. 5, 2 (2005).

    Article  Google Scholar 

  6. Peixoto, A., Monteiro, M., Rocha, B. & Veiga-Fernandes, H. Genome Res. 14, 1938–1947 (2004).

    Article  CAS  Google Scholar 

  7. Tsuji, K. et al. Cancer Res. 66, 303–306 (2006).

    Article  CAS  Google Scholar 

  8. Uehara, H., Osada, T. & Ikai, A. Ultramicroscopy 100, 197–201 (2004).

    Article  CAS  Google Scholar 

  9. Wagatsuma, A. et al. J. Exp. Biol. 208, 2389–2398 (2005).

    Article  CAS  Google Scholar 

  10. Bahcall, O.G. Mol. Syst. Biol. 1, 2005.0015 (2005).

    Article  Google Scholar 

  11. Cottingham, K. Anal. Chem. 76, 235A–238A (2004).

    CAS  PubMed  Google Scholar 

  12. Longo, D. & Hasty, J. Mol. Syst. Biol. 2, 64 (2006).

    Article  Google Scholar 

  13. Marcus, J.S., Anderson, W.F. & Quake, S.R. Anal. Chem. 78, 956–958 (2006).

    Article  CAS  Google Scholar 

  14. Hong, J.W., Studer, V., Hang, G., Anderson, W.F. & Quake, S.R. Nat. Biotechnol. 22, 435–439 (2004).

    Article  CAS  Google Scholar 

  15. Eberwine, J. Biotechniques 20, 584–591 (1996).

    CAS  PubMed  Google Scholar 

  16. Tang, F. et al. Nat. Methods 6, 377–382 (2009).

    Article  CAS  Google Scholar 

  17. Raineri, I., Moroni, C. & Senn, H.P. Nucleic Acids Res. 19, 4010 (1991).

    Article  CAS  Google Scholar 

  18. Roeder, T. Nucleic Acids Res. 26, 3451–3452 (1998).

    Article  CAS  Google Scholar 

  19. Fellmann, F., Pretet, J.L. & Fellmann, D. Biotechniques 21, 766–770 (1996).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank H. Tsunoda for his advice on cell culturing and single-cell sampling, K. Murakawa for his helpful suggestions, and M. Shirai for his advice on estimating the experimental errors.

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Authors and Affiliations

  1. Hitachi Central Research Laboratory, Tokyo, Japan

    Kiyomi Taniguchi, Tomoharu Kajiyama & Hideki Kambara

Authors
  1. Kiyomi Taniguchi
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  2. Tomoharu Kajiyama
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  3. Hideki Kambara
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Contributions

K.T. helped design the experiments, performed all the experiments, analyzed the data and helped prepare the manuscript. T.K. invented the method used for sample reuse and discussed the experimental result. H.K. helped design the experiments, analyzed the data and helped prepare the manuscript.

Corresponding author

Correspondence to Hideki Kambara.

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Supplementary Text and Figures

Supplementary Figures 1–4, Supplementary Tables 1–6, Supplementary Protocol, Supplementary Methods (PDF 519 kb)

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Taniguchi, K., Kajiyama, T. & Kambara, H. Quantitative analysis of gene expression in a single cell by qPCR. Nat Methods 6, 503–506 (2009). https://doi.org/10.1038/nmeth.1338

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