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.
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.
Consolandi
Clarissa
C
Institute of Biomedical Technologies, National Research Council, ITB-CNR, Via Fratelli Cervi 93, I-20090 Segrate, MI, Italy. clarissa.consolandi@itb.cnr.it
Severgnini
Marco
M
Frosini
Andrea
A
Caramenti
Giancarlo
G
De Fazio
Marco
M
Ferrara
Francesco
F
Zocco
Anna
A
Fischetti
Alessandra
A
Palmieri
Michele
M
De Bellis
Gianluca
G
D000458:Cyanobacteria / Q000235:genetics*; D004247:DNA / Q000032:analysis* / Q000097:blood / Q000378:metabolism; D004867:Equipment Design; D005798:Genes, Bacterial; D015894:Genome, Human; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D008904:Miniaturization; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D017655:Silicon Compounds / Q000737:chemistry*; D000514:alpha 1-Antichymotrypsin / Q000235:genetics*
D017655:Silicon Compounds; D000514:alpha 1-Antichymotrypsin; D004247:DNA