A droplet microfluidic approach to single-stream nucleic acid isolation and mutation detection.
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.
Department of Biomedical Engineering, Johns Hopkins University, Baltimore, Maryland, USA. Address: 122 Clark Hall, 3400 N Charles St, Baltimore, Maryland, USA.
Dong Jin
DJ
Shin
Department of Biomedical Engineering, Johns Hopkins University, Baltimore, Maryland, USA. Address: 122 Clark Hall, 3400 N Charles St, Baltimore, Maryland, USA.
Yi
Y
Zhang
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.
Tza-Huei
TH
Wang
Droplet PCR; Droplet microfluidics; Melting curve analysis; Mutation detection