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Microfluidic single-cell real-time PCR for comparative analysis of gene expression patterns

Sanches-Freire, V., Ebert, A.D., Kalisky, T., Quake, S.R., Wu, J.C.

Single-cell quantitative real-time PCR (qRT-PCR) combined with high-throughput arrays allows the analysis of gene expression profiles at a molecular level in approximately 11 h after cell sample collection. We present here a high-content microfluidic real-time platform as a powerful tool for comparatively investigating the regulation of developmental processes in single cells. This approach overcomes the limitations involving heterogeneous cell populations and sample amounts, and may shed light on differential regulation of gene expression in normal versus disease-related contexts. Furthermore, high-throughput single-cell qRT-PCR provides a standardized, comparative assay for in-depth analysis of the mechanisms underlying human pluripotent stem cell self-renewal and differentiation.

Citation

Sanches-Freire, V., Ebert, A.D., Kalisky, T., Quake, S.R., Wu, J.C. "Microfluidic single-cell real-time PCR for comparative analysis of gene expression patterns" Nature Protocols (2012): 829–38