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Virtual microfluidics for digital quantification and single-cell sequencing.

ABSTRACT: We have developed hydrogel-based virtual microfluidics as a simple and robust alternative to complex engineered microfluidic systems for the compartmentalization of nucleic acid amplification reactions. We applied in-gel digital multiple displacement amplification (dMDA) to purified DNA templates, cultured bacterial cells and human microbiome samples in the virtual microfluidics system, and demonstrated whole-genome sequencing of single-cell MDA products with excellent coverage uniformity and markedly reduced chimerism compared with products of liquid MDA reactions.

SUBMITTER: Xu L

PROVIDER: S-EPMC5007149 | biostudies-literature | 2016 Sep

REPOSITORIES: biostudies-literature

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Virtual microfluidics for digital quantification and single-cell sequencing.

Xu Liyi L Brito Ilana L IL Alm Eric J EJ Blainey Paul C PC

Nature methods 20160801 9

We have developed hydrogel-based virtual microfluidics as a simple and robust alternative to complex engineered microfluidic systems for the compartmentalization of nucleic acid amplification reactions. We applied in-gel digital multiple displacement amplification (dMDA) to purified DNA templates, cultured bacterial cells and human microbiome samples in the virtual microfluidics system, and demonstrated whole-genome sequencing of single-cell MDA products with excellent coverage uniformity and ma ...[more]

PMID: 27479330

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