A droplet microfluidic approach to single-stream nucleic acid isolation and mutation detection

Dong Jin Shin; Yi Zhang; Tza-Huei Wang

doi:10.1007/s10404-013-1305-7

A droplet microfluidic approach to single-stream nucleic acid isolation and mutation detection

Microfluid Nanofluidics. 2014 Aug;17(2):425-430. doi: 10.1007/s10404-013-1305-7.

Authors

Dong Jin Shin¹, Yi Zhang¹, Tza-Huei Wang²

Affiliations

¹ Department of Biomedical Engineering, Johns Hopkins University, Baltimore, Maryland, USA. Address: 122 Clark Hall, 3400 N Charles St, Baltimore, Maryland, USA.
² Department of Biomedical Engineering, Johns Hopkins University, Baltimore, Maryland, USA. Department of Mechanical Engineering, Johns Hopkins University, Baltimore, Maryland, USA. Institute of NanoBioTechnology, Baltimore, Maryland, USA. Sidney Kimmel Comprehensive Cancer Center, Baltimore, Maryland, USA. Address: 108 Latrobe Hall, 3400 N Charles St, Baltimore, Maryland, USA, Telephone: 410-516-7086.

Abstract

In this work, a droplet microfluidic platform for genetic mutation detection from crude biosample is described. Single-stream integration of nucleic acid isolation and amplification is realized on a simple fluidic cartridge. Subsequent DNA melting curve is employed with signal normalizing algorithm to differentiate heterozygous K-ras codon 12 c.25G>A mutant from the wildtype. This technique showcases an alternative to modular bench-top approaches for genetic mutation screening, which is of interest to decentralized diagnostic platforms.

Keywords: Droplet PCR; Droplet microfluidics; Melting curve analysis; Mutation detection.

Abstract

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