Pinched flow fractionation devices for detection of single nucleotide polymorphisms

Lab on a Chip
Asger Vig LarsenAnders Kristensen

Abstract

We demonstrate a new and flexible microfluidic based method for genotyping single nucleotide polymorphisms (SNPs). The method relies on size separation of selectively hybridized polystyrene microspheres in a microfluidic pinched flow fractionation (PFF) device. The microfluidic PFF devices with 13 mum deep channels were fabricated by thermal nanoimprint lithography (NIL) in a thin film of cyclic-olefin copolymer (mr-I T85) on a silicon wafer substrate, and the channels were sealed by thermal polymer bonding. Streptavidin coated polystyrene microspheres with a mean diameter of 3.09 microm and 5.6 microm were functionalized with biotin-labeled oligonucleotides for the detection of a mutant (Mt) or wild-type (Wt) DNA sequence in the HBB gene, respectively. Hybridization to functionalized beads was performed with fluorescent targets comprising synthetic DNA oligonucleotides or amplified RNA, synthesized using human DNA samples from individuals with point mutations in the HBB gene. Following a stringent wash, the beads were separated in a PFF device and the fluorescent signal from the beads was analyzed. Patients being wildtypes, heterozygotes or mutated respectively for the investigated mutation could reliably be diagnosed in the P...Continue Reading

References

Nov 10, 2001·Electrophoresis·H AnderssonG Stemme
Dec 6, 2001·Nature Reviews. Genetics·A C Syvänen
Mar 10, 2004·Lab on a Chip·Elisabeth Verpoorte
Aug 17, 2005·Clinica Chimica Acta; International Journal of Clinical Chemistry·Sherry A Dunbar

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Citations

Apr 24, 2010·Medical & Biological Engineering & Computing·Ali Asgar S BhagatChwee Teck Lim
Mar 1, 2012·Sensors·Rasim GuldikenJiang Zhe
Jul 17, 2012·Methods : a Companion to Methods in Enzymology·Julien AutebertJean-Louis Viovy
Mar 15, 2014·Langmuir : the ACS Journal of Surfaces and Colloids·Julien AutebertGovind V Kaigala
Oct 30, 2015·Lab on a Chip·Marie PødenphantRodolphe Marie
Jul 6, 2013·Biosensors & Bioelectronics·Yu-Tzu ChenJing-Tang Yang
Apr 4, 2015·Analytical Chemistry·Xinyu Lu, Xiangchun Xuan
Apr 15, 2010·Optics Express·Asger Laurburg VigAnders Kristensen
Oct 20, 2010·Lab on a Chip·Kyung Heon LeeHyung Jin Sung

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