Short title: Colorectal Cancer Subtype Biomarker Assay Analytical Validation of Multiplex Biomarker Assay to Stratify Colorectal Cancer Samples into Molecular Subtypes Chanthirika Ragulan 1,2,& , Katherine Eason 1,& , Elisa Fontana 1,2 , Gift Nyamundanda 1,2 , Yatish Patil 1,2 , Pawan Poudel 1 , Rita T. Lawlor 3,4 , Maguy Del Rio 5 , Koo Si-Lin 6 , Tan Wah Siew 7 , Francesco Sclafani 8 , Ruwaida Begum 8 , Larissa S. Teixeira Mendes 2 , Pierre Martineau 5 , Aldo Scarpa 3,4 , Iain Beehuat Tan 7,9,10 , David Cunningham 2,8 and Anguraj Sadanandam 1,2,* 1 Division of Molecular Pathology, The Institute of Cancer Research, London, United Kingdom 2 Centre for Molecular Pathology, The Royal Marsden NHS Foundation Trust, London, United Kingdom 3 ARC-Net Centre for Applied Research on Cancer, University and Hospital Trust of Verona, Verona, Italy 4 Department of Pathology and Diagnostics, University and Hospital Trust of Verona, Verona, Italy 5 Institut de Recherche en Cancérologie de Montpellier, Institut National de la Santé et de la Recherche Médicale, U896, Université Montpellier, Centre Régional de Lutte contre le Cancer Val d'Aurelle Paul Lamarque, Montpellier, France. 6 National Cancer Centre Singapore, Singapore 7 Singapore General Hospital, Singapore 8 Department of Medicine, The Royal Marsden NHS Foundation Trust, London, United Kingdom 9 Genome Institute of Singapore, Singapore 10 Duke-NUS Medical School, Singapore & These authors contributed equally to this work * Correspondence to Anguraj Sadanandam, Ph.D., Systems and Precision Cancer Medicine Team, Institute of Cancer Research (ICR), 15 Cotswold Road, Sutton, Surrey, SM2 5NG, United Kingdom; anguraj.sadanandam@icr.ac.uk Keywords: colorectal cancer subtypes, CRC subtypes, subtype diagnostic assay, biomarker prediction, multiplex biomarker assay, nCounter platform, mutations, class prediction, FFPE Article category: Research Article (Tumor Markers and Signatures) certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. The copyright holder for this preprint (which was not this version posted April 12, 2018. . https://doi.org/10.1101/174847 doi: bioRxiv preprint