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Published May 1, 2006 | Supplemental Material
Journal Article Open

Microfluidic Single-Cell mRNA Isolation and Analysis

Abstract

Single-cell gene expression analysis holds great promise for studying diverse biological systems, but methodology to process these precious samples in a reproducible, quantitative, and parallel fashion remains challenging. Here, we utilize microfluidics to isolate picogram and subpicogram mRNA templates, as well as to synthesize cDNA from these templates. We demonstrate single-cell mRNA isolation and cDNA synthesis, provide quantitative calibrations for each step in the process, and measure gene expression in individual cells. The techniques presented here form the foundation for highly parallel single-cell gene expression studies.

Additional Information

© 2006 American Chemical Society. Received for review October 31, 2005. Accepted February 24, 2006. We thank Kathy Burke for assistance with cell culture and real-time PCR. We also acknowledge Alejandra Torres for assistance with device fabrication and Carl Hansen and Sebastian Maerkl for helpful discussions. This work was supported by a National Research Service Award (T32GM07616) from the National Institute of General Medical Sciences (J.S.M.) and National Institutes of Health (NIH) grant NIH 1RO1 HG002644-01A1.

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Supplemental Material - ac0519460si20060213_014811.pdf

Supplemental Material - ac0519460si20060213_014928.pdf

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