Polymerase chain reaction of 2-kb cyanobacterial gene and human anti-α1-chymotrypsin gene from genomic DNA on the in-check single-use microfabricated silicon chip

被引:20
作者
Consolandi, C
Severgnini, M
Frosini, A
Caramenti, G
De Fazio, M
Ferrara, F
Zocco, A
Fischetti, A
Palmieri, M
De Bellis, G
机构
[1] CNR, ITB, Natl Res Council, I-20090 Segrate, MI, Italy
[2] STMicroelect, Microfluid Div, I-20041 Agrate Brianza, Italy
关键词
polymerase chain reaction; single-use silicon chip; genomic DNA; miniaturization;
D O I
10.1016/j.ab.2006.03.019
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The microfabricated chip is a promising format for automating and miniaturizing the multiple steps of genotyping. We tested an innovative silicon biochip (In-Check Lab-on-Chip; STMicroelectronics, Agrate Brianza, Italy) designed for polymerase chain reaction (PCR) analysis of complex biological samples. The chip is mounted on a 1 x 3-in(2). plastic slide that provides the necessary mechanical, thermal, electrical, and fluidic connections. A temperature control system drives the chip to the desired temperatures, and a graphical user interface allows experimenters to define cycling conditions and monitor reactions in real time. During thermal cycling, we recorded a cooling rate of 3.2 degrees C/s and a heating rate of 11 degrees C/s. The temperature maintained at each thermal plateau was within 0.13 C of the programmed temperature at three sensors. From 0.5 ng/mu l genomic DNA, the In-Check device successfully amplified the 2060-by cyanobacterial 16S rRNA gene and the 330-bp human anti-alpha(1)-chymotrypsin gene. The shortest PCR protocol that produced an amplicon by capillary electrophoresis comprised 30 cycles and was 22.5 min long. These thermal cycling characteristics suggest that the In-Check device will permit future development of a genotyping lab-on-a-chip device, yielding results in a short time from a limited amount of biological starting material. 2006 Elsevier Inc. (c) All rights reserved.
引用
收藏
页码:191 / 197
页数:7
相关论文
共 19 条
[1]   Development of a universal microarray based on the ligation detection reaction and 16S rRNA gene polymorphism to target diversity of cyanobacteria [J].
Castiglioni, B ;
Rizzi, E ;
Frosini, A ;
Sivonen, K ;
Rajaniemi, P ;
Rantala, A ;
Mugnai, MA ;
Ventura, S ;
Wilmotte, A ;
Boutte, C ;
Grubisic, S ;
Balthasart, P ;
Consolandi, C ;
Bordoni, R ;
Mezzelani, A ;
Battaglia, C ;
De Bellis, G .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2004, 70 (12) :7161-7172
[2]   Electrokinetically synchronized polymerase chain reaction microchip fabricated in polycarbonate [J].
Chen, JF ;
Wabuyele, M ;
Chen, HW ;
Patterson, D ;
Hupert, M ;
Shadpour, H ;
Nikitopoulos, D ;
Soper, SA .
ANALYTICAL CHEMISTRY, 2005, 77 (02) :658-666
[3]   Chip PCR .2. Investigation of different PCR amplification systems in microfabricated silicon-glass chips [J].
Cheng, J ;
Shoffner, MA ;
Hvichia, GE ;
Kricka, LJ ;
Wilding, P .
NUCLEIC ACIDS RESEARCH, 1996, 24 (02) :380-385
[4]   Comparison of chemical cleaning methods of glass in preparation for silanization [J].
Cras, JJ ;
Rowe-Taitt, CA ;
Nivens, DA ;
Ligler, FS .
BIOSENSORS & BIOELECTRONICS, 1999, 14 (8-9) :683-688
[5]   Integrated system for rapid PCR-based DNA analysis in microfluidic devices [J].
Khandurina, J ;
McKnight, TE ;
Jacobson, SC ;
Waters, LC ;
Foote, RS ;
Ramsey, JM .
ANALYTICAL CHEMISTRY, 2000, 72 (13) :2995-3000
[6]   Chemical amplification: Continuous-flow PCR on a chip [J].
Kopp, MU ;
de Mello, AJ ;
Manz, A .
SCIENCE, 1998, 280 (5366) :1046-1048
[7]   Selective Surface Modification in Silicon Microfluidic Channels for Micromanipulation of Biological Macromolecules [J].
Li, Youli ;
Pfohl, Thomas ;
Kim, Joon Heon ;
Yassa, Mario ;
Wen, Zhiyu ;
Kim, Mahn Won ;
Safinya, Cyrus R. .
BIOMEDICAL MICRODEVICES, 2001, 3 (03) :239-244
[8]   A new promoter polymorphism in the alpha-1-antichymotrypsin gene is a disease modifier of Alzheimer's disease [J].
Licastro, F ;
Chiappelli, M ;
Grimaldi, LME ;
Morgan, K ;
Kalsheker, N ;
Calabrese, E ;
Ritchie, A ;
Porcellini, E ;
Salani, G ;
Franceschi, M ;
Canal, N .
NEUROBIOLOGY OF AGING, 2005, 26 (04) :449-453
[9]   Self-contained, fully integrated biochip for sample preparation, polymerase chain reaction amplification, and DNA microarray detection [J].
Liu, RH ;
Yang, JN ;
Lenigk, R ;
Bonanno, J ;
Grodzinski, P .
ANALYTICAL CHEMISTRY, 2004, 76 (07) :1824-1831
[10]   DNA amplification and hybridization assays in integrated plastic monolithic devices [J].
Liu, YJ ;
Rauch, CB ;
Stevens, RL ;
Lenigk, R ;
Yang, JN ;
Rhine, DB ;
Grodzinski, P .
ANALYTICAL CHEMISTRY, 2002, 74 (13) :3063-3070