title: Patterns of cross-contamination in a multispecies population genomic project: detection, quantification, impact, and solutions.
abstract: Contamination is a well-known but often neglected problem in molecular biology. Here, we investigated the prevalence of cross-contamination among 446 samples from 116 distinct species of animals, which were processed in the same laboratory and subjected to subcontracted transcriptome sequencing.
  
  
  
  Using cytochrome oxidase 1 as a barcode, we identified a minimum of 782 events of between-species contamination, with approximately 80% of our samples being affected. An analysis of laboratory metadata revealed a strong effect of the sequencing center: nearly all the detected events of between-species contamination involved species that were sent the same day to the same company. We introduce new methods to address the amount of within-species, between-individual contamination, and to correct for this problem when calling genotypes from base read counts.
  
  
  
  We report evidence for pervasive within-species contamination in this data set, and show that classical population genomic statistics, such as synonymous diversity, the ratio of non-synonymous to synonymous diversity, inbreeding coefficient FIT, and Tajima's D, are sensitive to this problem to various extents. Control analyses suggest that our published results are probably robust to the problem of contamination. Recommendations on how to prevent or avoid contamination in large-scale population genomics/molecular ecology are provided based on this analysis.
authors.affiliation: UMR5554 - Institute of Evolutionary Sciences, University Montpellier, CNRS, IRD, EPHE, Place Eugène Bataillon, CC64, 34095, Montpellier, France.|UMR7144 - Adaptation et Diversité en Milieu Marin - CNRS, Université Pierre et MarieCurie, Station Biologique de Roscoff, 29680, Roscoff, France.
authors.forename: Marion
authors.initials: M
authors.lastname: Ballenghien
authors.affiliation: UMR5554 - Institute of Evolutionary Sciences, University Montpellier, CNRS, IRD, EPHE, Place Eugène Bataillon, CC64, 34095, Montpellier, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Faivre
authors.affiliation: UMR5554 - Institute of Evolutionary Sciences, University Montpellier, CNRS, IRD, EPHE, Place Eugène Bataillon, CC64, 34095, Montpellier, France. nicolas.galtier@univ-montp2.fr.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Galtier
mesh_terms: D059018:DNA Contamination; D030541:Databases, Genetic; D003576:Electron Transport Complex IV / Q000235:genetics; D005828:Genetics, Population; D023281:Genomics; D056186:Metagenomics; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D011336:Probability; D013045:Species Specificity
chemical_list: D003576:Electron Transport Complex IV
keywords: Animals; Genotyping; RNAseq; SNP calling; Transcriptome; Within-species