Microfluidics for Army Synthetic Biology—DNA ENTRAP: A Microfluidic Tool for Enhanced DNA Transfer

Author(s):

Jose A. Wippold, Monica Chu, Rebecca Renberg, Margaret Hurley, and Bryn Adams

Abstract:

Droplet microfluidic platforms have been broadly used to facilitate DNA transfer in mammalian and bacterial hosts via methods such as transformation, transfection, and conjugation, as introduced in our previous work. Herein, we recapitulate our method for conjugal DNA transfer between Bacillus subtilis strains in a droplet for increased conjugation efficiency and throughput of an otherwise laborious protocol. By co-incubating the donor and recipient strains in a droplet, our method confines cells into close proximity and allows for increased cell-to-cell interactions. This methodology is advantageous because it has the potential to automate and accelerate the genetic modification of undomesticated organisms that may be difficult to cultivate. This device is also designed for modularity and can be integrated into a variety of experimental workflows in which fine tuning of donor-to-recipient cell ratios, growth rates, and media substrate concentrations may be necessary.