Quantitative and qualitative analyses of DNA fragments based on electrical charge detection on a portable electrophoresis device.
A concept regarding DNA fragments electrophoretic analyses by directly detecting electrical charges is proposed. The arrival time and voltage of charged DNA fragments with different charge-to-mass ratio could be detected using the custom-made micro electronic circuits. These time and voltage information imply the size and intensity information acquired from the conventional slab gel image by fluorescent labeling. A prototype of the portable electrophoresis device consists of a flow channel with the dimension of 35 mm (length) x 0.5 mm (width) x 0.2 mm (depth) on an acrylic substrate, and the detection circuit with amplification gain of 10,000 and analogous filter bandwidth between 0.1 Hz and 10 Hz has been developed. A simple experiment was carried out to demonstrate the feasibility of proposed idea. The volume of 2mul of the DNA ladder (1 Kb Plus DNA ladder, Invitrogen, U.S.A.) with the diluted concentration of 0.1mug/mul was loaded into the reservoir when applying the electrical field of 12.5 V/cm to both end of the flow channel, which was only filled with TBE solution. The preliminary results showed that the developed electrophoresis device can pick up the electrical signals of un-separated DNA fragments with total mass of 0.2 mug , and the magnitude is 0.6 V . Micro flow channels fabricated by an excimer-laser machine and low-noise amplifier with high gain, e.g. 100,000 are being processed. Moreover, HEC (hydroxyethylcellulose) solution will be utilized as the media in the micro channels for DNA fragments separation.
Chen
Gin-Shin
GS
Div. of Med. Eng. Res., Nat. Health Res. Inst., Taipei, Taiwan.
Chen
Sheng-Fu
SF
Lu
Chih-Cheng
CC