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Exploiting biased reptation for continuous flow preparative DNA fractionation in a versatile microfluidic platform.

ABSTRACT: A new approach is presented for preparative, continuous flow fractionation of sub-10-kbp DNA fragments, which exploits the variation in the field-dependent mobility of the DNA molecules based on their length. Orthogonally pulsed electric fields of significantly different magnitudes are applied to a microchip filled with a sieving matrix of 1.2% agarose gel. Using this method, we demonstrate a high-resolution separation of 0.5, 1, 2, 5, and 10 kbp DNA fragments within 2 min. During the separation, DNA fragments are also purified from other ionic species. Preparative fractionation of sub-10-kbp DNA molecules plays an important role in second-generation sequencing. The presented device performs rapid high-resolution fractionation and it can be reliably manufactured with simple microfabrication procedures.

SUBMITTER: Gumuscu B

PROVIDER: S-EPMC6444976 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature

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Exploiting biased reptation for continuous flow preparative DNA fractionation in a versatile microfluidic platform.

Gumuscu Burcu B Bomer Johan G JG de Boer Hans L HL van den Berg Albert A Eijkel Jan C T JCT

Microsystems & nanoengineering 20170522

A new approach is presented for preparative, continuous flow fractionation of sub-10-kbp DNA fragments, which exploits the variation in the field-dependent mobility of the DNA molecules based on their length. Orthogonally pulsed electric fields of significantly different magnitudes are applied to a microchip filled with a sieving matrix of 1.2% agarose gel. Using this method, we demonstrate a high-resolution separation of 0.5, 1, 2, 5, and 10 kbp DNA fragments within 2 min. During the separation ...[more]

PMID: 31057856

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