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Robust label-free microRNA detection using one million ISFET array

  • Anurup Ganguli
  • Yoshihiko Watanabe
  • Michael T. Hwang
  • Jui-Cheng Huang
  • Rashid Bashir
Article

Abstract

Detection of nucleic acid molecules is one of the most pervasive assays in biology, medicine, and agriculture applications. Currently, most comely used DNA/RNA detection platforms use fluorescence labeling and require lab-scale setting for performing the assay. There is a need for developing less expensive, label-free, and rapid detection of biomolecules with minimal utilization of resources. Use of electrical approaches for detection of biomolecules by utilizing their inherent charge is a promising direction for biosensing assays. Here, we report a 1024 × 1024 array of Ion Sensitive Field Effect Transistors (ISFET) as label free sensors for detection of nucleic acid molecules. Using PNA probe functionalized on these ISFET array, we robustly detected miRNA Let-7b by measuring changes in drain current after hybridization of target molecules with concentration as low as 1 nM. We demonstrate that mismatched or non-complementary target molecules resulted in statistically smaller changes. Most importantly, the high-density sensor array shows unprecedented reliability and robustness with P values <0.0001 for all experiments. Practical implementation of this platform could have a wide range of applications in high-throughput nucleic acid genotyping, detection of amplified pathogenic nucleic acid, detection of cell-free DNA, and electrical readouts for current hybridization-based DNA biomolecular assays.

Keywords

Electrical DNA sensor Electrical microarray Multiplexed biosensing 1024 × 1024 array ISFET 

Notes

Acknowledgements

We thank the staff at the Micro and Nanotechnology Laboratory at UIUC for facilitating the chip fabrication. The work was funded by NSF grant 1534126 and the University of Illinois at Urbana-Champaign.

Author contributions

A.G., Y.W. and R.B conceived the idea and designed the study. A.G. and Y.W., designed and performed the experiments. J.H. assisted with the experiments and provided intellectual inputs. M.H., A.G., Y.W. and R.B wrote and edited the manuscript.

Compliance with ethical standards

Competing interest

The authors declare no competing financial interests.

Supplementary material

10544_2018_290_MOESM1_ESM.docx (2.7 mb)
Fig. S1 (DOCX 2803 kb)

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Anurup Ganguli
    • 1
    • 2
  • Yoshihiko Watanabe
    • 2
  • Michael T. Hwang
    • 2
  • Jui-Cheng Huang
    • 3
  • Rashid Bashir
    • 1
    • 2
    • 4
  1. 1.Department of BioengineeringUniversity of Illinois at Urbana-ChampaignUrbanaUSA
  2. 2.Micro and Nanotechnology LaboratoryUniversity of Illinois at Urbana-ChampaignUrbanaUSA
  3. 3.Taiwan Semiconductor Manufacturing CompanyTaiwan,Republic of China
  4. 4.Carle Illinois College of MedicineChampaignUSA

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