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Biomedical Microdevices

, Volume 16, Issue 5, pp 661–670 | Cite as

A polymer Lab-on-a-Chip for genetic analysis using the arrayed primer extension on microarray chips

  • Simone L. MarassoEmail author
  • Domenico Mombello
  • Matteo Cocuzza
  • Davide Casalena
  • Ivan Ferrante
  • Alessandro Nesca
  • Piret Poiklik
  • Kadri Rekker
  • Anu Aaspollu
  • Sergio Ferrero
  • Candido F. Pirri
Article

Abstract

In this work a polymer lab-on-a-chip (LOC), fabricated through MEMS technology, was employed to execute a genetic protocol for the Single Nucleotide Polymorphisms (SNPs) detection. The LOC was made in Poly (methyl methacrylate) (PMMA) and has two levels: the lower one for the insertion and mixing of the reagents, the upper one for the interfacing with the DNA microarray chip. The hereditary hearing loss was chosen as case of study, since the demand for testing such a particular disorder is high and genetics behind the condition is quite clear. The Arrayed Primer EXtension (APEX) genetic protocol was implemented on the LOC to analyze the SNPs. A low density (for detection of up to 10 mutations) and a high density microarray chips (for detection of 245 mutations in 12 genes), containing the primers for the extension, were employed to carry out the APEX reaction on the LOC. Both the microarray chips provide a signal to noise ratio and efficiency comparable with a detection obtained in a conventional protocol in standard conditions. Moreover, significant reduction of the employed PCR volume (from 30 μL to 10 μL) was obtained using the low density chip.

Keywords

Lab-on-a-Chip DNA microarray PCR (polymerase chain reaction) APEX (Arrayed Primer EXtension) Single Nucleotide Polymorphisms (SNPs) 

Notes

Acknowledgments

This work was funded and accomplished in the framework of the MANUNET ERA-Net Project “AUDAX - Automation of a Device based on APEX technology” (2008 Manunet Transnational Call), in cooperation with project EU31699 funded by Enterprise Estonia (LOC (Lab-on-a-chip) based genotyping technology –AUDAX applied research/product development.

Supplementary material

10544_2014_9869_MOESM1_ESM.docx (84 kb)
Online Resource 1 (DOCX 83 kb)

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Simone L. Marasso
    • 1
    • 2
    Email author
  • Domenico Mombello
    • 1
  • Matteo Cocuzza
    • 1
    • 2
  • Davide Casalena
    • 1
  • Ivan Ferrante
    • 1
  • Alessandro Nesca
    • 3
  • Piret Poiklik
    • 4
  • Kadri Rekker
    • 4
  • Anu Aaspollu
    • 4
  • Sergio Ferrero
    • 1
    • 3
  • Candido F. Pirri
    • 1
  1. 1.Xlab – Materials and Microsystems Laboratory, Applied Science and Technology Department (DISAT)Polytechnic of Turin – Latemar UnitChivasso (Turin)Italy
  2. 2.IMEM-CNRParmaItaly
  3. 3.Microla SRLChivasso (TO)Italy
  4. 4.Asper BiotechTartuEstonia

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