Handmade microfluidic device for biochemical applications in emulsion.

Area of Science:

• Biochemistry
• Molecular Biology
• Biotechnology

Background:

• Uniform droplet generation is crucial for reproducible biochemical reactions.
• Existing methods can be expensive or require specialized equipment.
• High-throughput screening demands precise control over reaction volumes and conditions.

Purpose of the Study:

• To develop a simple, inexpensive flow-focusing device for uniform droplet generation.
• To demonstrate the device's utility in synthesizing and analyzing biomolecules.
• To assess the device's impact on quantitative analysis in high-throughput screening.

Main Methods:

• Fabrication of a flow-focusing device using commercially available components.
• Synthesis of a ligase ribozyme (bcI 23) using T7 RNA polymerase on magnetic microbeads within emulsion droplets.
• Expression of a lambda Cro protein derivative using an Escherichia coli cell-free system in emulsion.
• Detection of synthesized RNA and protein using flow cytometry.

Main Results:

• Uniform emulsion droplets were successfully generated for biochemical reactions.
• Class I ligase ribozyme (bcI 23) was synthesized and ligated on microbeads.
• A lambda Cro protein derivative was expressed in an *E. coli* cell-free system within emulsion.
• The flow-focusing device significantly reduced the coefficient of variation for RNA and protein display.

Conclusions:

• The developed flow-focusing device is a cost-effective tool for generating uniform droplets.
• The device facilitates efficient synthesis and quantitative analysis of RNA and proteins.
• This technology is advantageous for high-throughput screening applications in synthetic biology and drug discovery.