Journal article

A new normalization for Nanostring nCounter gene expression data

Ramyar Molania, Johann A Gagnon-Bartsch, Alexander Dobrovic, Terence P Speed

Nucleic Acids Research | OXFORD UNIV PRESS | Published : 2019


The Nanostring nCounter gene expression assay uses molecular barcodes and single molecule imaging to detect and count hundreds of unique transcripts in a single reaction. These counts need to be normalized to adjust for the amount of sample, variations in assay efficiency and other factors. Most users adopt the normalization approach described in the nSolver analysis software, which involves background correction based on the observed values of negative control probes, a within-sample normalization using the observed values of positive control probes and normalization across samples using reference (housekeeping) genes. Here we present a new normalization method, Removing Unwanted Variation-..

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Awarded by National Health and Medical Research Council of Australia

Funding Acknowledgements

National Health and Medical Research Council of Australia Program Grant [1054618 to T.P.S.]; Cancer Australia for the Lung Cancer DNA Repair Study (to A. D.); The University of Melbourne international Australian postgraduate scholarships (MIRS and MIFRS) (to R.M.); Cancer Therapeutics CRC CTx PhD Top Up Scholarship (to R.M.). Funding for open access charge: Walter & Eliza Hall Institute and NHMRC Grant.