DNA analysis on microfabricated electrophoretic devices with bubble cells.
Microfluidic devices with bubble cells have been fabricated on poly(methyl methacrylate) (PMMA) plates and have been employed for the analysis of DNA using polyethylene oxide (PEO) solutions. First, the separation channel was fabricated using a wire-imprinting method. Then, wires with greater sizes or a razor blade glued in a polycarbonate plate was used to fabricate bubble cells, with sizes of 190-650 microm. The improvements in resolution and sensitivity have been achieved for large DNA (> 603 base pair, bp) using such devices, which depend on the geometry of the bubble cell. The main contributor for optimal resolution is mainly due to DNA migration at lower electric field strengths inside the bubble cell. On the other hand, slight losses of resolution for small DNA fragments have been found mainly due to diffusion, supported by the loss of resolution when separating two small solutes. With a bubble cell of 75 microm (width) x 500 microm (depth), the sensitivity improvement up to 17-fold has been achieved for the 271 bp fragment in the separation of PhiX-174/HaeIII DNA restriction fragments. We have also found that a microfluidic device with a bubble cell of 360 microm x 360 microm is appropriate for DNA analysis. Such a device has been used for separating DNA ranging from 8 to 2176 bp and polymerase chain reaction (PCR) products amplified after 30 cycles, with rapidity and improvements in the sensitivity as well as resolution.
Tseng
Wei-Lung
WL
Department of Chemistry, National Taiwan University, Taipei, Taiwan, ROC.
Lin
Yang-Wei
YW
Chen
Ko-Chun
KC
Chang
Huan-Tsung
HT
D001483:Base Sequence; D004247:DNA / Q000032:analysis* / Q000235:genetics / Q000302:isolation & purification; D004586:Electrophoresis / Q000295:instrumentation* / Q000379:methods / Q000706:statistics & numerical data; D006801:Humans; D008297:Male; D008832:Microchemistry / Q000295:instrumentation; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity
D004247:DNA