High-throughput cell DNA screening using Digital PCR

High-throughput cell DNA screening using Digital PCR

The use of water-in-oil droplets in microfluidics in high-throughput screening is rapidly gaining acceptance. The main application areas currently involve screening cells as well as genetic material for various mutations or activity. Here the aim is to isolate single DNA molecules and analyze the enzymes and proteins resulting from their expression.

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High-throughput cell encapsulation-graph

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There were several steps to the study:

  • First, two separate emulsions were generated. The first one encapsulated an aqueous-phase of PCR and DNA mixture in an organic continuous phase on a dedicated chip.

The second emulsion was generated encapsulating an aqueous-phase IVT (In Vitro Transmission) mixture in the same organic phase.

  • Lastly, both emulsions were then fused one droplet at a time to control which DNA sequence is translated at the end of the process.

For these studies, it is very important that the droplets are generated at the correct frequency, and at uniform size. That’s where the use of the MFCS™-EZ is critical with the generation of very stable flows and the accurate control of the different phases.

A graphic representation of the droplet chip used to combine the PCR and IVT droplets is below.

The MFCS™-EZ, Fluigent pressure controller, controlled by software, allows to create your two emulsions. The Flow Units allows one to control the flow-rates during the experiment.