The family Trichomycteridae is one of the most diverse groups of freshwater catfishes in South and Central America with eight subfamilies, 41 genera and more than 300 valid species. Its members are widely distributed throughout South America, reaching Costa Rica in Central America and are recognized by extraordinary anatomical specializations and trophic diversity. In order to assess the phylogenetic relationships of Trichomycteridae, we collected sequence data from ultraconserved elements (UCEs) of the genome from 141 specimens of Trichomycteridae and 12 outgroup species. We used a concatenated matrix to assess the phylogenetic relationships by Bayesian inference (BI) and maximum likelihood (ML) searches and a coalescent analysis of species trees. The results show a highly resolved phylogeny with broad agreement among the three distinct analyses, providing overwhelming support for the monophyletic status of subfamily Trichomycterinae including Ituglanis and Scleronema. Previous relationship hypotheses among subfamilies are strongly corroborated, such as the sister relationship between Copionodontinae and Trichogeninae forming a sister clade to the remaining trichomycterids and the intrafamilial clade TSVSG (Tridentinae-Stegophilinae-Vandelliinae-Sarcoglanidinae-Glanapteryginae). Monophyly of Glanapteryginae and Sarcoglanidinae was not supported and the enigmatic Potamoglanis is placed outside tridentinae. Unraveling the relationships of major sections of the Tree of Life is one of the most daunting challenges of the evolutionary biology. Massively parallel DNA sequencing (so-called Next-gen sequencing) is a promising tool that is helping to resolve the interrelationships of longstanding problematic taxa 1-4. One of the most common classes of phylogenomic methods involves the sequence capture of nuclear regions in the flanks and cores of the ultraconserved elements (UCEs) 2. The more variable flanking UCE regions allow a better resolution of nodes across a broad range of evolutionary timescales in a given phylogeny 2. As variation in the flanks increases with distance from the core UCE, this combined approach displays a balance between having a high enough substitution rate while minimizing saturation, thus providing information for estimating phylogenies at multiple evolutionary timescales 2,3. Recent studies of actinopterygians 5 , flatfishes 6 , cichlids 7 , ostariophysan 8 , acanthomorphs 9 , Loricariidae 10 , knifefishes 11 , among other vertebrates groups 3,12 , have shown that UCEs are excellent markers for phylogenetic studies because of their ubiquity among taxonomic groups 13 , low degrees of paralogy 14 , and low saturation 3. According to Gilbert et al. 15 , the phylogenetic informativeness of the combined flank and core regions of UCEs outperfoms protein-coding genes used in multilocus studies. Additionally, phylogenomic approaches are characterized by their potential to collect data from at least one order of magnitude more loci than the traditional sequencing techniques applied to protein-coding le...