Consolandi Clarissa, Severgnini Marco, Frosini Andrea, Caramenti Giancarlo, De Fazio Marco, Ferrara Francesco, Zocco Anna, Fischetti Alessandra, Palmieri Michele, De Bellis Gianluca
Institute of Biomedical Technologies, National Research Council, ITB-CNR, Via Fratelli Cervi 93, I-20090 Segrate, MI, Italy.
Anal Biochem. 2006 Jun 15;353(2):191-7. doi: 10.1016/j.ab.2006.03.019. Epub 2006 Mar 31.
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 1x3-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 degrees C of the programmed temperature at three sensors. From 0.5 ng/microl genomic DNA, the In-Check device successfully amplified the 2060-bp 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.
微制造芯片是一种很有前景的形式,可实现基因分型多个步骤的自动化和小型化。我们测试了一种创新的硅生物芯片(In-Check芯片实验室;意法半导体公司,意大利阿格拉特布里安扎),该芯片专为对复杂生物样本进行聚合酶链反应(PCR)分析而设计。芯片安装在一块1×3英寸(2)的塑料载玻片上,该载玻片提供必要的机械、热、电和流体连接。温度控制系统将芯片驱动至所需温度,图形用户界面允许实验人员定义循环条件并实时监测反应。在热循环过程中,我们记录到冷却速率为3.2℃/秒,加热速率为11℃/秒。在三个传感器处,每个热平台维持的温度与设定温度的偏差在0.13℃以内。从0.5 ng/μl基因组DNA开始,In-Check设备成功扩增了2060 bp的蓝藻16S rRNA基因和330 bp的人抗α(1)-糜蛋白酶基因。通过毛细管电泳产生扩增子的最短PCR方案包括30个循环,时长为22.5分钟。这些热循环特性表明,In-Check设备将有助于未来基因分型芯片实验室设备的开发,能在短时间内从有限量的生物起始材料中得出结果。