title: A MEMS-Based Approach to Single Nucleotide Polymorphism Genotyping.
abstract: Genotyping of single nucleotide polymorphisms (SNPs) allows diagnosis of human genetic disorders associated with single base mutations. Conventional SNP genotyping methods are capable of providing either accurate or high-throughput detection, but are still labor-, time-, and resource-intensive. Microfluidics has been applied to SNP detection to provide fast, low-cost, and automated alternatives, although these applications are still limited by either accuracy or throughput issues. To address this challenge, we present a MEMS-based SNP genotyping approach that uses solid-phase-based reactions in a single microchamber on a temperature control chip. Polymerase chain reaction (PCR), allele specific single base extension (SBE), and desalting on microbeads are performed in the microchamber, which is coupled with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) to analyze the SBE product. Experimental results from genotyping of the SNP on exon 1 of the HBB gene, which causes sickle cell anemia, demonstrate the potential of the device for rapid, accurate, multiplexed and high-throughput detection of SNPs.
authors.affiliation: Department of Mechanical Engineering, Columbia University, New York, NY, USA.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhu
authors.affiliation: Department of Mechanical Engineering, Columbia University, New York, NY, USA ; Department of Chemical Engineering, Columbia University, New York, NY, USA.
authors.forename: Mirkó
authors.initials: M
authors.lastname: Palla
authors.affiliation: Department of Mechanical Engineering, Columbia University, New York, NY, USA ; Department of Mechanical and Industrial Engineering, University of Brescia, Brescia, BS, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Ronca
authors.affiliation: Department of Obstetrics and Gynecology, Columbia University, New York, NY, USA.
authors.forename: Ronald
authors.initials: R
authors.lastname: Warpner
authors.affiliation: Department of Chemical Engineering, Columbia University, New York, NY, USA.
authors.forename: Jingyue
authors.initials: J
authors.lastname: Ju
authors.affiliation: Department of Mechanical Engineering, Columbia University, New York, NY, USA.
authors.forename: Qiao
authors.initials: Q
authors.lastname: Lin
keywords: Bead-Based Polymerase Chain Reaction; Bead-Based Single Base Extension; Genotyping; Microfluidics; Single Nucleotide Polymorphisms
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