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Direct multiplexed measurement of gene expression with color-coded probe pairs

Geiss, Gary K. and Bumgarner, Roger E. and Birditt, Brian and Dahl, Timothy and Dowidar, Naeem and Dunaway, Dwayne L. and Fell, H. Perry and Ferree, Sean and George, Renee D. and Grogan, Tammy and James, Jeffrey J. and Maysuria, Malini and Mitton, Jeffrey D. and Oliveri, Paola and Osborn, Jennifer L. and Peng, Tao and Ratcliffe, Amber L. and Webster, Philippa J. and Davidson, Eric H. and Hood, Leroy and Dimitrov, Krassen (2008) Direct multiplexed measurement of gene expression with color-coded probe pairs. Nature Biotechnology, 26 (3). pp. 317-325. ISSN 1087-0156.

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We describe a technology, the NanoString nCounter gene expression system, which captures and counts individual mRNA transcripts. Advantages over existing platforms include direct measurement of mRNA expression levels without enzymatic reactions or bias, sensitivity coupled with high multiplex capability, and digital readout. Experiments performed on 509 human genes yielded a replicate correlation coefficient of 0.999, a detection limit between 0.1 fM and 0.5 fM, and a linear dynamic range of over 500-fold. Comparison of the NanoString nCounter gene expression system with microarrays and TaqMan PCR demonstrated that the nCounter system is more sensitive than microarrays and similar in sensitivity to real-time PCR. Finally, a comparison of transcript levels for 21 genes across seven samples measured by the nCounter system and SYBR Green real-time PCR demonstrated similar patterns of gene expression at all transcript levels.

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Hood, Leroy0000-0001-7158-3678
Additional Information:© 2008 Macmillan Publishers Limited. Received 17 September 2007; Accepted 28 January 2008; Published online 17 February 2008; Corrected 16 May 2008. We would like to acknowledge Edward Kelly, at the Center for DNA Sequencing and Gene Analysis, UW School of Pharmacy, Department of Pharmaceutics for TaqMan analysis and the University of Washington's Center for Array Technologies for processing of Affymetrix arrays. Poliovirus (PV) stocks were the kind gift of Kurt Gustin's laboratory (University of Idaho). Author contributions: G.K.G. designed the experiments, analyzed data and prepared the manuscript. D.L.D., S.F., P.J.W. and G.K.G. designed and developed the NanoString system described. T.D. and J.D.M. developed probe selection and image analysis software, respectively. N.D. performed all nCounter assays described. B.B., R.D.G., T.G., J.J.J., M.M., J.L.O. and A.L.R. provided ideas and technical support. T.P. and P.O. provided microarray and real-time PCR data and samples. R.E.B. contributed to experimental design, scientific direction and manuscript preparation. H.P.F., E.H.D. and L.H. provided scientific direction and experimental concepts. K.D. provided the ideas and design of this technology and participated in the initial research. he authors declare competing financial interests. The following authors are current or past employees of and/or have an equity stake in NanoString technologies: G.K.G., B.B., T.D., N.D., D.L.D., H.P.F., S.F., R.D.G., T.G., J.J.J., M.M., J.D.M., J.L.O., A.L.R., P.J.W., E.H.D., L.H. and K.D.
Errata:Corrigendum: Direct multiplexed measurement of gene expression with color-coded probe pairs Gary K Geiss, Roger E Bumgarner, Brian Birditt, Timothy Dahl, Naeem Dowidar, Dwayne L Dunaway, H Perry Fell, Sean Ferree, Renee D George, Tammy Grogan, Jeffrey J James, Malini Maysuria, Jeffrey D Mitton, Paola Oliveri, Jennifer L Osborn, Tao Peng, Amber L Ratcliffe, Philippa J Webster, Eric H Davidson & Leroy Hood Corrigenda Nature Biotechnology 26, 709 (2008) doi:10.1038/nbt0608-709c
Issue or Number:3
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:64398
Deposited By: George Porter
Deposited On:19 Feb 2016 19:20
Last Modified:29 Oct 2019 22:02

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