We are developing a new technique to insert foreign DNA into a living cell using a microelectromechanical system. This new technique relies on electrical forces to move DNA in a nonuniform electric field. To better understand this phenomenon, we perform integrated modeling and experiments of DNA electrophoresis. This paper describes the protocol and presents the results for DNA motion experiments using fabricated gel electrophoresis devices. We show that DNA motion is strongly correlated with ion transport (current flow) in the system. A better understanding of electrophoretic fundamentals allows for the creation of a mathematical model to predict the motion of DNA during electrophoresis in both uniform and nonuniform electric fields. The mathematical model is validated within 4% through comparison with the experimental results.
This paper describes the protocol and presents the results for DNA motion experiments using fabricated macroscale gel electrophoresis devices. Gel electrophoresis is a process used to separate/move DNA, RNA or protein molecules using an electric field through a gel matrix (electrolytic solution). In electrolytic solutions, the current conduction is due to a transport of ions (anions and cations). A better understanding of electrophoretic fundamentals allows for modeling the motion of DNA during electrophoresis. The model is validated through comparison with the experimental results. The model and experimental validation will be used to improve the process of cellular nanoinjection of DNA, currently in development in our lab.
This paper reports the effects of various parameters on the attraction and repulsion of DNA to and from a silicon lance. An understanding of DNA motion is crucial for a new approach to insert DNA, or other foreign microscopic matter, into a living cell. The approach, called nanoinjection, uses electrical forces to attract and repel the desired substance to a micromachined lance designed to pierce the cell membranes. We have developed mathematical models to predict the trajectory of DNA. The mathematical model allows investigation of the attraction/repulsion process by varying specific parameters. We find that the ground electrode placement, lance orientation and lance penetration significantly affect attraction or repulsion efficiency, while the gap, lance direction, lance tip width, lance tip half-angle and lance tip height do not.
Electrophoretic systems commonly use metal electrodes in their construction. This paper explores and reports the differences in the electrophoretic motion of DNA (decomposition voltage, electrical field, etc.) when one electrode is constructed from a semiconductor, silicon, rather than metal. Experimental VI (voltage-current) curves for different electrode configurations (using steel and silicon) are presented. Experimental results are used to update and validate the mathematical model to reflect the differences in material selection. In addition, the model predicts large curved-field motion for DNA motion. The model helps to quantify the effect of parameters on DNA motion in biological microelectromechanical systems in order to improve device designs and protocols.
We are developing a new technique, called nanoinjection, to insert foreign DNA into a living cell. Such DNA transfection is commonly used to create transgenic organisms vital to the study of genetics, immunology, and many other biological sciences. In nanoinjection, DNA, which has a net negative charge, is electrostatically attracted to a micromachined lance. The lance then pierces the cell membranes, and the voltage on the lance is reversed, repelling the DNA into the cell. This paper presents a mathematical model to predict the motion (trajectory) of DNA particles within a cell in the presence of the electric field developed by the lance and the substrate. The model is used to predict the scattering of DNA through the cell due to electrostatic repulsion. We are currently preparing experiments which will be used to validate the model.
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