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Issue 22, 2012
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Microfluidic extraction, stretching and analysis of human chromosomal DNA from single cells

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Abstract

We describe a microfluidic device for the extraction, purification and stretching of human chromosomal DNA from single cells. A two-dimensional array of micropillars in a microfluidic polydimethylsiloxane channel was designed to capture a single human cell. Megabase-long DNA strands released from the cell upon lysis are trapped in the micropillar array and stretched under optimal hydrodynamic flow conditions. Intact chromosomal DNA is entangled in the array, while other cellular components are washed from the channel. To demonstrate the entrapment principle, a single chromosome was hybridized to whole chromosome paints, and imaged by fluorescence microscopy. DNA extracted from a single cell and small cell populations (less than 100) was released from the device by restriction endonuclease digestion under continuous flow and collected for off-chip analysis. Quantification of the extracted material reveals that the microdevice efficiently extracts essentially all chromosomal DNA. The device described represents a novel platform to perform a variety of analyses on chromosomal DNA at the single cell level.

Graphical abstract: Microfluidic extraction, stretching and analysis of human chromosomal DNA from single cells

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Publication details

The article was received on 21 Aug 2012, accepted on 10 Sep 2012 and first published on 10 Sep 2012


Article type: Paper
DOI: 10.1039/C2LC40955K
Citation: Lab Chip, 2012,12, 4848-4854
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    Microfluidic extraction, stretching and analysis of human chromosomal DNA from single cells

    J. J. Benítez, J. Topolancik, H. C. Tian, C. B. Wallin, D. R. Latulippe, K. Szeto, P. J. Murphy, B. R. Cipriany, S. L. Levy, P. D. Soloway and H. G. Craighead, Lab Chip, 2012, 12, 4848
    DOI: 10.1039/C2LC40955K

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