Identificação de Loci microssatélites em Viola - Loricariichthys anus (Valenciennes, 1840).

Abstract

n Brazil there is a rich diversity of fish, however, little is known about the biology of different species. The Viola - Loricariichthys anus - is a native species of southern South America which has great economic potential and requires great attention towards conservation. Microsatellite markers are a great tool for the genetic study, conservation and breeding of fish, especially with the next-generation sequencing technologies that reduce costs and optimize time and results. The objective was to identify microsatellite loci for the non-template species L. anus and know how the species is and behave genetically. DNA extraction was performed by the modified glass silica extraction protocol. A genomic library shotgun paired-end was prepared by Kit Nextera Illumina DNA Library protocol and then sequenced in a MiSeq Illumina sequencer and 1,103,580 of obtained readings were analyzed in PAL_FINDER_V0.02.03 program to recognize the microsatellite markers. Were obtained 444 microsatellite loci potentially amplifiable (PALs), of which 161 were dinucleotides, 65 were trinucleotides, 170 were tetranucleotides, 32 and 16 pentanucleotides and hexanucleotides. A group of twelve loci were chosen for validation by PCR. These 6 were amplified - four dinucleotides and two tetranucleotídes. However, we can define the sequencing strategy through library shotgun paired-end MiSeq platform (Illumina) presented itself effective even with a low number of PALs when compared with other studies, as well as being quick and inexpensive to develop microsatellite markers for non-template species such as the viola.