Droplet-based microfluidic systems for high-throughput single DNA molecule isothermal amplification and analysis
- Resource Type
- Authors
- Agnes Janoshazi; J. Brian Hutchison; Valérie Taly; Linas Mazutis; Benjamin J. Miller; Jean-Christophe Baret; Lucas Frenz; Ali Fallah Araghi; Oliver J. Miller; Andrew D. Griffiths; Michael Ryckelynck; Darren Link
- Source
- Analytical Chemistry
Analytical Chemistry, American Chemical Society, 2009, 81 (12), pp.4813-21. ⟨10.1021/ac900403z⟩
Analytical Chemistry, 2009, 81 (12), pp.4813-21. ⟨10.1021/ac900403z⟩
- Subject
- endocrine system
Microfluidics
Loop-mediated isothermal amplification
Analytical chemistry
02 engineering and technology
01 natural sciences
complex mixtures
Analytical Chemistry
MESH: Nucleic Acid Amplification Techniques
chemistry.chemical_compound
Molecule
Throughput (business)
Fusion
Translation system
010401 analytical chemistry
technology, industry, and agriculture
MESH: DNA
Temperature
DNA
Microfluidic Analytical Techniques
021001 nanoscience & nanotechnology
Dna amplification
eye diseases
MESH: Temperature
Intercalating Agents
0104 chemical sciences
MESH: Intercalating Agents
chemistry
MESH: Microfluidic Analytical Techniques
Biophysics
0210 nano-technology
Nucleic Acid Amplification Techniques
- Language
- ISSN
- 1520-6882
0003-2700
International audience; We have developed a method for high-throughput isothermal amplification of single DNA molecules in a droplet-based microfluidic system. DNA amplification in droplets was analyzed using an intercalating fluorochrome, allowing fast and accurate "digital" quantification of the template DNA based on the Poisson distribution of DNA molecules in droplets. The clonal amplified DNA in each 2 pL droplet was further analyzed by measuring the enzymatic activity of the encoded proteins after fusion with a 15 pL droplet containing an in vitro translation system.