Microfluidic devices and methods of use in the formation and control of nanoreactors

A method for preparing nucleic acids for sequencing, the method comprising:

• providing a first dispersed phase solution comprising primer-bound microbeads and primer pairs and a second dispersed phase solution comprising nucleic acid templates via respective first and second inlet channels in a coalescence module of a microfluidic device;
• coalescing by the coalescence module droplets of the first and the second dispersed phase solution to generate nanoreactor droplets, a plurality of the droplets each comprising one of the primer-bound microbeads, at least one of the nucleic acid templates, and a label for amplified DNA;
• amplifying the nucleic acids templates with the primer-bound microbeads and the primer pairs to generate bead-bound amplified nucleic acids;
• separating using the label the droplets into a first population of the droplets that each contain the amplified DNA and a second population of the droplets that do not contain the amplified DNA;
• breaking the first population of the droplets;
• depositing the bead-bound amplified nucleic acids onto a solid support for sequencing.