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1.
Fresenius J Anal Chem ; 371(2): 190-4, 2001 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-11678190

RESUMEN

Over the last 50 years or so Traditional Chinese medicine (TCM) has been subject to intensive basic and clinical research. Although the effectiveness and remarkable safety of TCM have been documented after controlled clinical studies, there are several herbal and animal parts that are toxic or difficult to identify. DNA polymorphism-based assays have recently been developed for the identification of herbal medicines. In this approach, small amounts of DNA are amplified by the polymerase chain reaction and the reactions products are analyzed by gel electrophoresis, sequencing, or hybridization with species-specific probes. With the DNA based identification of TCM materials as an example, chip-based analytical micro devices were developed with the goal of fabricating an integrated device that will enable sample preparation, amplification, and analysis on a single microchip-based device ("lab-on-a-chip"). The application of a silicon-based polymerase chain reaction microreactor and a DNA microarray for the DNA sequence-based identification of toxic medicinal plants is reported here.


Asunto(s)
Medicina Tradicional China , Análisis de Secuencia por Matrices de Oligonucleótidos/métodos , Plantas Medicinales/genética , ADN de Plantas/análisis , Diseño de Equipo , Humanos , Microquímica , Análisis de Secuencia por Matrices de Oligonucleótidos/instrumentación , Plantas Medicinales/clasificación , Reacción en Cadena de la Polimerasa/métodos , Silicio
2.
Anal Chem ; 72(17): 4242-7, 2000 Sep 01.
Artículo en Inglés | MEDLINE | ID: mdl-10994990

RESUMEN

A Si-based miniaturized device for the polymerase chain reaction (PCR) has been developed. The device has Pt temperature sensors and heaters integrated on top of the reaction chamber for real-time accurate temperature sensing and control. Reaction temperature of the device is digitally controlled to achieve a temperature accuracy of +/-0.025 degrees C. The effects of PCR protocol optimization on the amplification performance of the surface-passivated chip reactor have been investigated in detail and quantitatively compared with that of the conventional thermal cycler. In this study, four traditional Chinese medicine genes including Fritillaria cirrhosa, Cartharmus tinctorius, Adenophora lobophilla, and Stephania tetrandra are used as model template. With appropriate chamber surface treatment (chlorotrimethylsilane/polyadenylic acid or SiO2 coatings), the device demonstrates amplification as efficient as that in the conventional thermal cycler at optimized MgCl2 concentration. The amplified DNA has concentration higher than 27 ng/microL, which is sufficient for subsequent on-chip analyses and detection. Experimental results reveal the importance of inclusion of BSA for an efficient amplification in the SiO2-passivated device and the excellent reusability of the device with a simple cleaning step.


Asunto(s)
ADN de Plantas/análisis , Medicina Tradicional China , Reacción en Cadena de la Polimerasa , Temperatura
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