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Microfluidic Single-Cell mRNA Isolation and Analysis

Marcus, Joshua S. and Anderson, W. French and Quake, Stephen R. (2006) Microfluidic Single-Cell mRNA Isolation and Analysis. Analytical Chemistry, 78 (9). pp. 3084-3089. ISSN 0003-2700. doi:10.1021/ac0519460. https://resolver.caltech.edu/CaltechAUTHORS:20161103-122919491

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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.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1021/ac0519460DOIArticle
http://pubs.acs.org/doi/full/10.1021/ac0519460PublisherArticle
http://pubs.acs.org/doi/suppl/10.1021/ac0519460Related ItemSupporting Information
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.
Funders:
Funding AgencyGrant Number
NIH Predoctoral FellowshipT32GM07616
National Institute of General Medical SciencesUNSPECIFIED
NIH1RO1 HG002644-01A1
Issue or Number:9
DOI:10.1021/ac0519460
Record Number:CaltechAUTHORS:20161103-122919491
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20161103-122919491
Official Citation:Microfluidic Single-Cell mRNA Isolation and Analysis Joshua S. Marcus, W. French Anderson, and, and Stephen R. Quake Analytical Chemistry 2006 78 (9), 3084-3089 DOI: 10.1021/ac0519460
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:71709
Collection:CaltechAUTHORS
Deposited By: Ruth Sustaita
Deposited On:03 Nov 2016 23:59
Last Modified:11 Nov 2021 04:49

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