| Autor: |
Raisi, Fariba, Blizard, Benjamin A., Raissi Shabari, Akbar, Ching, Jesus, Kintz, Gregory J., Mitchell, Jim, Lemoff, Asuncion, Taylor, Mike T., Weir, Fred, Western, Linda, Wong, Wendy, Joshi, Rekha, Howland, Pamela, Chauhan, Avinash, Nguyen, Peter, Petersen, Kurt E. |
| Zdroj: |
Journal of Separation Science (JSS) (formerly Journal of High Resolution Chromatography); March 2004, Vol. 27 Issue: 4 p275-283, 9p |
| Abstrakt: |
The growing importance of analyzing the human genome to detect hereditary and infectious diseases associated with specific DNA sequences has motivated us to develop automated devices to integrate sample preparation, real‐time PCR, and microchannel electrophoresis (MCE). In this report, we present results from an optimized compact system capable of processing a raw sample of blood, extracting the DNA, and performing a multiplexed PCR reaction. Finally, an innovative electrophoretic separation was performed on the post‐PCR products using a unique MCE system. The sample preparation system extracted and lysed white blood cells (WBC) from whole blood, producing DNA of sufficient quantity and quality for a polymerase chain reaction (PCR). Separation of multiple amplicons was achieved in a microfabricated channel 30 μm × 100 μm in cross section and 85 mm in length filled with a replaceable methyl cellulose matrix operated under denaturing conditions at 50°C. By incorporating fluorescent‐labeled primers in the PCR, the amplicons were identified by a two‐color (multiplexed) fluorescence detection system. Two base‐pair resolution of single‐stranded DNA (PCR products) was achieved. We believe that this integrated system provides a unique solution for DNA analysis. |
| Databáze: |
Supplemental Index |
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