Modification of a poly(methyl methacrylate) injection-molded microchip and its use for high performance analysis of DNA.
Inexpensive and permanently modified poly(methyl methacrylate)(PMMA) microchips were fabricated by an injection-molding process. A novel sealing method for plastic microchips at room temperature was introduced. Run-to-run and chip-to-chip reproducibility was good, with relative standard deviation values between 1-3% for the run-to-run and less than 2.1% for the chip-to-chip comparisons. Acrylonitrile-butadiene-styrene (ABS) was used as an additive in PMMA substrates. The proportions of PMMA and ABS were optimized. ABS may be considered as a modifier, which obviously improved some characteristics of the microchip, such as the hydrophilicity and the electro-osmotic flow (EOF). The detection limit of Rhodamine 6G dye for the modified microchip on the home-made microchip analyzer showed a dramatic 100-fold improvement over that for the unmodified PMMA chip. A detection limit of the order of 10(-20) mole has been achieved for each injected psiX-174/HaeIII DNA fragment with the baseline separation between 271 and 281 bp, and fast separation of 11 DNA restriction fragments within 180 seconds. Analysis of a PCR product from the tobacco ACT gene was performed on the modified microchip as an application example.
Zhou
Xiao-Mian
XM
Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, PR China.
Dai
Zhong-Peng
ZP
Liu
Xin
X
Luo
Yong
Y
Wang
Hui
H
Lin
Bing-Cheng
BC
D004247:DNA / Q000032:analysis*; D008855:Microscopy, Electron, Scanning; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation*; D016133:Polymerase Chain Reaction; D019904:Polymethyl Methacrylate / Q000737:chemistry*; D015203:Reproducibility of Results; D012235:Rhodamines
D012235:Rhodamines; D004247:DNA; D019904:Polymethyl Methacrylate