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Anchored multiplex amplification on a microelectronic chip array

Nature Biotechnology volume 18, pages 199–204 (2000)Cite this article

Abstract

We have developed a method for anchored amplification on a microchip array that allows amplification and detection of multiple targets in an open format. Electronic anchoring of sets of amplification primers in distinct areas on the microchip permitted primer-primer interactions to be reduced and distinct zones of amplification created, thereby increasing the efficiency of the multiplex amplification reactions. We found strand displacement amplification (SDA) to be ideal for use in our microelectronic chip system because of the isothermal nature of the assay, which provides a rapid amplification system readily compatible with simple instrumentation. Anchored SDA supported multiplex DNA or RNA amplification without decreases in amplification efficiency. This microelectronic chip-based amplification system allows multiplexed amplification and detection to be performed on the same platform, streamlining development of any nucleic acid-based assay.

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Figure 1: Schematic representation of anchored strand displacement amplification (SDA).
Figure 2: Anchored SDA amplification of human Factor V DNA.
Figure 3: 10-plex anchored SDA amplification.
Figure 4: Anchored genomic DNA amplification.
Figure 5: Anchored RT-SDA.

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Acknowledgements

The authors would like to gratefully acknowledge the intellectual contributions and support of Drs. Terry Walker, Dan MacLaurin, and Doug Malinowski, and Cathy Spargo of Becton-Dickinson, and Drs. Tina S. Nova, Jim P. O'Connell, Michael Heller, John Carrino, and Prashant Mehta of Nanogen, Inc. throughout the course of this work. The authors are especially grateful to Drs. Ray Radtke, Lana Feng, and Geoff Landis, and Dana Vollmer of Nanogen, Inc. for their contributions in support of permeation layer, primer design, and SDA in earlier stages of this work. Supported under Award number 97-LB-VX-0004 from the Office of Justice Programs, National Institute of Justice, Department of Justice. Points of view in this document are those of the authors and do not necessarily represent the official position of the US Department of Justice.

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Authors and Affiliations

  1. Department of Molecular Biology, Nanogen, Inc., San Diego, CA

    Lorelei Westin, Carolyn Miller, Ling Wang & Michael Nerenberg

  2. Department of Genomics, Nanogen, Inc., San Diego, CA

    Xiao Xu

  3. Department of Genomics, Nanotronics, Inc., San Diego, CA

    Carl F. Edman

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  1. Lorelei Westin
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Correspondence to Lorelei Westin.

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Westin, L., Xu, X., Miller, C. et al. Anchored multiplex amplification on a microelectronic chip array. Nat Biotechnol 18, 199–204 (2000). https://doi.org/10.1038/72658

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