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Nat Methods. 2016 Sep;13(9):759-62. doi: 10.1038/nmeth.3955. Epub 2016 Aug 1.

Virtual microfluidics for digital quantification and single-cell sequencing.

Xu L1,2, Brito IL1,2,3, Alm EJ1,2,3, Blainey PC1,2.

Author information

1
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
2
The Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
3
The Center for Microbiome Informatics and Therapeutics, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.

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.

PMID:
27479330
PMCID:
PMC5007149
DOI:
10.1038/nmeth.3955
[Indexed for MEDLINE]
Free PMC Article

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