Development of PCR conditions in a silicon microreactor DNA-amplification device
A silicon microsystem was developed which functions as a miniaturised DNA-amplification device. The system represents a technology platform for performing a polymerase chain reaction (PCR) with reduced volumes of 7 µL. The silicon microreactor was fabricated using silicon bulk micromachining, and a...
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| Published in: | International journal of environmental analytical chemistry Vol. 84; no. 11; pp. 821 - 833 |
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| Main Authors: | , , , , , |
| Format: | Journal Article |
| Language: | English |
| Published: |
Taylor & Francis Group
15-09-2004
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| Subjects: | |
| Online Access: | Get full text |
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| Summary: | A silicon microsystem was developed which functions as a miniaturised DNA-amplification device. The system represents a technology platform for performing a polymerase chain reaction (PCR) with reduced volumes of 7 µL. The silicon microreactor was fabricated using silicon bulk micromachining, and a platinum heater was fabricated on a Pyrex substrate. A miniaturised DNA-amplification system permitted rapid heating and cooling, and shorter reaction times of 30 min were achieved. In this work, biocompatibility issues are addressed; conditions for efficient PCR in a silicon-based microreactor are established for the amplification of 500 bp DNA from the Escherichia coli bacteriophage Lambda; and the conditions are verified by amplifying a 255 bp region from the Mycobacterium tuberculosis rpoB gene. This work describes the PCR volume scale down experiments that were conducted and concentrations of the reactants; Taq polymerase, oligonucleotides, MgCl
2
and template DNA were determined for DNA-amplification reactions with this novel device. |
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| Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
| ISSN: | 0306-7319 1029-0397 |
| DOI: | 10.1080/0306731042000268143 |