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Nucleic Acids Research, 2001, Vol. 29, No. 23 e118
© 2001 Oxford University Press

Signal amplification by rolling circle amplification on DNA microarrays

Girish Nallur*, Chenghua Luo, Linhua Fang, Stephanie Cooley, Varshal Dave, Jeremy Lambert, Kari Kukanskis, Stephen Kingsmore, Roger Lasken and Barry Schweitzer

Molecular Staging Inc., 300 George Street, New Haven, CT 06511, USA

While microarrays hold considerable promise in large-scale biology on account of their massively parallel analytical nature, there is a need for compatible signal amplification procedures to increase sensitivity without loss of multiplexing. Rolling circle amplification (RCA) is a molecular amplification method with the unique property of product localization. This report describes the application of RCA signal amplification for multiplexed, direct detection and quantitation of nucleic acid targets on planar glass and gel-coated microarrays. As few as 150 molecules bound to the surface of microarrays can be detected using RCA. Because of the linear kinetics of RCA, nucleic acid target molecules may be measured with a dynamic range of four orders of magnitude. Consequently, RCA is a promising technology for the direct measurement of nucleic acids on microarrays without the need for a potentially biasing preamplification step.

* To whom correspondence should be addressed. Tel: +1 203 772 5018; Fax; +1 203 776 5278; Email: girishn{at}molecularstaging.com Present addresses: Varshal Dave, Axon Instruments, Arlington, TX 76012, USA Jeremy Lambert, HTS BioSystems, Hopkinton, MA 01748, USA


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