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1.
J Sep Sci ; 28(3): 225-33, 2005 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-15776923

RESUMO

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.


Assuntos
DNA/análise , Análise de Sequência com Séries de Oligonucleotídeos/instrumentação , Polimetil Metacrilato/química , Microscopia Eletrônica de Varredura , Reação em Cadeia da Polimerase , Reprodutibilidade dos Testes , Rodaminas
2.
Se Pu ; 20(2): 156-8, 2002 Mar.
Artigo em Chinês | MEDLINE | ID: mdl-12541975

RESUMO

UV labeling detection has been commonly used to determine the association constants between lectins and saccharides, but the interaction is always between the labeled carbohydrates, rather than the truly underivatized carbohydrates, and lectins. In order to directly detect saccharides during the study on the interaction of glucose and its derivatives with lectins (e.g., concanavalin A), a capillary zone electrophoretic method with detection at a wavelength of 195 nm has been developed. The influences of various separation conditions including buffer concentration, pH and voltage were investigated. By using an uncoated silica capillary (50 microns i.d., 375 microns o.d., 48.5 cm of total length, and 44.0 cm to the detector) and 50 mmol/L Na2HPO(4)-50 mmol/L NaH2PO4 solution (near to the physiological pH of 7.4) as buffer, the underivatized sugars, including glucosamine, N-acetylglucosamine, glucose, and sodium gluconate, were sufficiently separated within 11 min at an applied voltage of 10 kV. On-column UV monitoring allowed the detection of these compounds at less than 4 mmol/L level, and quantification by the peak area method allowed reproducible determination of them at least at their respective concentration ranges. The method is characterized by its simplicity, rapidity, and reproducibility, and should be useful for the analysis of the interaction of glucose and its derivatives with lectins.


Assuntos
Eletroforese Capilar/métodos , Glucose/análise , Lectinas/análise , Concanavalina A/análise , Concanavalina A/isolamento & purificação , Gluconatos/análise , Gluconatos/isolamento & purificação , Glucose/análogos & derivados , Glucose/isolamento & purificação , Lectinas/isolamento & purificação , Espectrofotometria Ultravioleta
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