Nucleic acid and protein extraction from electropermeabilized E. coli cells on a microfluidic chip

Area of Science:

• Biotechnology
• Molecular Biology
• Microfluidics

Background:

• Bacterial nucleic acid and protein analysis requires efficient extraction methods.
• Existing protocols can be time-consuming and complex.
• There is a need for rapid, simple extraction of plasmid DNA, RNA, and reporter proteins like GFP.

Purpose of the Study:

• To develop a simple and rapid electropermeability technique for extracting biomolecules from E. coli.
• To enable simultaneous extraction of nucleic acids and proteins.
• To achieve cell survival during extraction using controlled voltage.

Main Methods:

• Utilized a microfluidic chip with integrated gold electrodes for electropermeability.
• Applied low DC voltages (below 0.5 V) to transiently permeabilize E. coli cell envelopes.
• Controlled voltage to achieve partial/transient opening for survival or complete lysis for DNA recovery.

Main Results:

• Successfully extracted RNA and Green Fluorescent Protein (GFP) using electropermeability at low voltages.
• Achieved plasmid DNA recovery by increasing voltage to 2 V, inducing complete cell lysis.
• Demonstrated that low voltages (below 0.5 V) allow transient pore formation, cell survival, and biomolecule release.

Conclusions:

• Electropermeability offers a rapid and versatile method for bacterial biomolecule extraction.
• The technique allows for controlled cell permeabilization, enabling either cell survival or lysis.
• This method provides a simple alternative for extracting RNA, proteins, and plasmid DNA from bacterial samples.