Polymerase chain reaction of 2-kb cyanobacterial gene and human anti-alpha1-chymotrypsin gene from genomic DNA on the In-Check single-use microfabricated silicon chip

Anal Biochem. 2006 Jun 15;353(2):191-7. doi: 10.1016/j.ab.2006.03.019. Epub 2006 Mar 31.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cyanobacteria / genetics*
  • DNA / analysis*
  • DNA / blood
  • DNA / metabolism
  • Equipment Design
  • Genes, Bacterial
  • Genome, Human
  • Humans
  • Microfluidic Analytical Techniques / instrumentation*
  • Miniaturization
  • Polymerase Chain Reaction / instrumentation
  • Polymerase Chain Reaction / methods*
  • Silicon Compounds / chemistry*
  • alpha 1-Antichymotrypsin / genetics*

Substances

  • Silicon Compounds
  • alpha 1-Antichymotrypsin
  • DNA