共 19 条
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
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机构: CNR, ITB, Natl Res Council, I-20090 Segrate, MI, Italy

Severgnini, M
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h-index: 0
机构: CNR, ITB, Natl Res Council, I-20090 Segrate, MI, Italy

Frosini, A
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h-index: 0
机构: CNR, ITB, Natl Res Council, I-20090 Segrate, MI, Italy

Caramenti, G
论文数: 0 引用数: 0
h-index: 0
机构: CNR, ITB, Natl Res Council, I-20090 Segrate, MI, Italy

De Fazio, M
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h-index: 0
机构: CNR, ITB, Natl Res Council, I-20090 Segrate, MI, Italy

Ferrara, F
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h-index: 0
机构: CNR, ITB, Natl Res Council, I-20090 Segrate, MI, Italy

Zocco, A
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h-index: 0
机构: CNR, ITB, Natl Res Council, I-20090 Segrate, MI, Italy

Fischetti, A
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h-index: 0
机构: CNR, ITB, Natl Res Council, I-20090 Segrate, MI, Italy

Palmieri, M
论文数: 0 引用数: 0
h-index: 0
机构: CNR, ITB, Natl Res Council, I-20090 Segrate, MI, Italy

De Bellis, G
论文数: 0 引用数: 0
h-index: 0
机构: CNR, ITB, Natl Res Council, I-20090 Segrate, MI, Italy
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.
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页码:191 / 197
页数:7
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