Bulk-micromachined submicroliter-volume PCR chip with very rapid thermal response and low power consumption.
The current paper describes the design, fabrication, and testing of a micromachined submicroliter-volume polymerase chain reaction (PCR) chip with a fast thermal response and very low power consumption. The chip consists of a bulk-micromachined Si component and hot-embossed poly(methyl methacrylate)(PMMA) component. The Si component contains an integral microheater and temperature sensor on a thermally well-isolated membrane, while the PMMA component contains a submicroliter-volume PCR chamber, valves, and channels. The micro hot membrane under the submicroliter-volume chamber is a silicon oxide/silicon nitride/silicon oxide (O/N/O) diaphragm with a thickness of 1.9 microm, resulting in a very low thermal mass. In experiments, the proposed chip only required 45 mW to heat the reaction chamber to 92 degrees C, the denaturation temperature of DNA, plus the heating and cooling rates are about 80 degrees C s(-1) and 60 degrees C s(-1), respectively. We validated, from the fluorescence results from DNA stained with SYBR Green I, that the proposed chip amplified the DNA from vector clone, containing tumor suppressor gene BRCA 1 (127 base pairs at 11th exon), after 30 thermal cycles of 3 s, 5 s, and 5 s at 92 degrees C, 55 degrees C, and 72 degrees C, respectively, in a 200 nL-volume chamber. As for specificity of DNA products, owing to difficulty in analyzing the very small volume PCR results from the micro chip, we vicariously employed the larger volume PCR products after cycling with the same sustaining temperatures as with the micro chip but with much slower ramping rates (3.3 degrees C s(-1) when rising, 2.5 degrees C s(-1) when cooling) within circa 20 minutes on a commercial PCR machine and confirmed the specificity to BRCA 1 (127 base pairs) with agarose gel electrophoresis. Accordingly, the fabricated micro chip demonstrated a very low power consumption and rapid thermal response, both of which are crucial to the development of a fully integrated and battery-powered instrument for a lab-on-a-chip DNA analysis.
Lee
Dae-Sik
DS
BioMEMS Team, ETRI, P.O. Box 106, Yuseong-Gu, Daejeon 305-350, Korea.
Park
Se Ho
SH
Yang
Haesik
H
Chung
Kwang-Hyo
KH
Yoon
Tae Hwan
TH
Kim
Sung-Jin
SJ
Kim
Kyuwon
K
Kim
Youn Tae
YT
D002138:Calibration; D004247:DNA / Q000032:analysis; D011211:Electric Power Supplies; D004867:Equipment Design; D006358:Hot Temperature; D008832:Microchemistry / Q000295:instrumentation*; D016133:Polymerase Chain Reaction / Q000295:instrumentation* / Q000379:methods*; D019904:Polymethyl Methacrylate / Q000737:chemistry; D015203:Reproducibility of Results; D017655:Silicon Compounds / Q000737:chemistry; D013696:Temperature
D017655:Silicon Compounds; D004247:DNA; D019904:Polymethyl Methacrylate