Genotyping of single nucleotide polymorphisms by melting curve analysis using thin film semi-transparent heaters integrated in a lab-on-foil system
2013 (English)In: Lab on a Chip, ISSN 1473-0197, E-ISSN 1473-0189, Vol. 13, no 11, 2075-2082 p.Article in journal (Refereed) Published
The recent technological advances in micro/nanotechnology present new opportunities to combine microfluidics with microarray technology for the development of small, sensitive, single-use, point-of-care molecular diagnostic devices. As such, the integration of microarray and plastic microfluidic systems is an attractive low-cost alternative to glass based microarray systems. This paper presents the integration of a DNA microarray and an all-polymer microfluidic foil system with integrated thin film heaters, which demonstrate DNA analysis based on melting curve analysis (MCA). A novel micro-heater concept using semi-transparent copper heaters manufactured by roll-to-roll and lift-off on polyethylene naphthalate (PEN) foil has been developed. Using a mesh structure, heater surfaces have been realized in only one single metallization step, providing more efficient and homogenous heating characteristics than conventional meander heaters. A robust DNA microarray spotting protocol was adapted on Parylene C coated heater-foils, using co-polymer poly(DMA-NAS-MAPS) to enable covalent immobilization of DNA. The heaters were integrated in a microfluidic channel using lamination foils and MCA of the spotted DNA duplexes showed single based discrimination of mismatched over matched target DNA-probes. Finally, as a proof of principle, we perform MCA on PCR products to detect the Leu7Pro polymorphism of the neutropeptide Y related to increased risk of Type II diabetes, BMI and depression.
Place, publisher, year, edition, pages
2013. Vol. 13, no 11, 2075-2082 p.
article, body mass, depression, devices, DNA determination, DNA microarray, film, foil, heating, melting curve analysis, non insulin dependent diabetes mellitus, nucleic acid analysis, polymerase chain reaction, priority journal, single nucleotide polymorphism, thin film heater
Biochemistry and Molecular Biology
IdentifiersURN: urn:nbn:se:kth:diva-123643DOI: 10.1039/c3lc50171jISI: 000318514400008ScopusID: 2-s2.0-84877682690OAI: oai:DiVA.org:kth-123643DiVA: diva2:628568
FunderScience for Life Laboratory - a national resource center for high-throughput molecular bioscience
QC 201306142013-06-142013-06-132013-06-14Bibliographically approved