RLeave: an in silico cross-validation protocol for transcript differential expression analysis. Authors: Matheus Costa e Silva 1 , Norma Lucena-Silva 2 , Juliana Doblas Massaro 1* , Eduardo Antônio Donadi 1* . * These authors contributed equally as senior authors. Institutions: 1 Division of Clinical Immunology, Department of Medicine, Ribeirão Preto Medical School, University of São Paulo, 14048-900, Ribeirão Preto, SP, Brazil. 2 Laboratory of Immunogenetics, Department of Immunology, Aggeu Magalhães Institute, 50740-465, Recife, PE, Brazil. Abstract Background and Objective: The massive parallel sequencing technology facilitates new discoveries in terms of transcript differential analysis; however, all the new findings must be validated, since the diversity of transcript expression may impair the identification of the most relevant ones. Methods: The proposed RLeave algorithm (implemented in the R environment) utilizes a combination of conventional analysis (classic edgeR) together with other mathematical methods (Leave-one-out sample technique and Decision Trees validation) to identify more relevant candidates to be in vitro or in silico validated.