Patagonian toothfish (Dissostichus eleginoides, Smith 1898) is a bentopelagic fish, from the Antarctic circumpolar, of high economic value. It has a long life, around 50 years and a large size and weight, around 2 meters long and 200 kilograms. Chile is the main exporter of Patagonian toohfish worldwide. At present the condition of this fish is exhausted, data of Subsecretaria de Pesca y Acuicultura, Chile, 2018. The De Novo assembly was made using the Trinity. To reduce gene redundancy, gene clustering was performed, 233,424 unigenes were detected. After assembly, the subsequent annotation with Blast was identified for functional annotation, in total 37,446 unigenes were recorded in different databases (NR, Swiss-Prot, Uniprot, KEGG and GO). A new analysis was performed by aligning the reads against the reference transcriptome for a deeper analysis. In this reanalysis, 223,405 single nucleotide polymorphism (SNP) markers were found on average per tissue. A total of 71,107 microsatellites (simple sequence repeats – SSR) were detected. The transcriptome is a very useful tool to begin understand the biology of an organism, especially if we want to start growing Patagonian toothfish. It is a long-lived fish that must understand the machinery necessary for its development and early production.