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Microfluidic long DNA sample preparation from cells

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Abstract

A number of outstanding problems in genomics, such as identifying structural variations and sequencing through centromeres and telomeres, stand poised to benefit tremendously from emerging long-read genomics technologies such as nanopore sequencing and genome mapping in nanochannels. However, optimal application of these new genomics technologies requires facile methods for extracting long DNA from cells. These sample preparation tools should be amenable to automation and minimize fragmentation of the long DNA molecules by shear. We present one such approach in a poly(dimethylsiloxane) device, where gel-based high molecular weight DNA extraction and continuous flow purification in a 3D cell culture-inspired geometry is followed by electrophoretic extraction of the long DNA from the miniaturized gel. Molecular combing reveals that the device produces molecules that are typically in excess of 100 kilobase pairs in size, with the longest molecule extending up to 4 megabase pairs. The microfluidic format reduces the standard day-long and labor-intensive DNA extraction process to 4 hours, making it a promising prototype platform for routine long DNA sample preparation.

Graphical abstract: Microfluidic long DNA sample preparation from cells

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Publication details

The article was received on 27 Oct 2018, accepted on 30 Nov 2018 and first published on 03 Dec 2018


Article type: Paper
DOI: 10.1039/C8LC01163J
Citation: Lab Chip, 2019, Advance Article
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    Microfluidic long DNA sample preparation from cells

    P. Agrawal and K. D. Dorfman, Lab Chip, 2019, Advance Article , DOI: 10.1039/C8LC01163J

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