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Quantitative analysis of gene expression in a single cell by qPCR

Nature Methods volume 6, pages 503–506 (2009)Cite this article

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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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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