Evaluation of eight protocols for genomic DNA extraction of Hypostomus commersoni (Valenciennes, 1836), Siluriformes: Loricariidae.
Priscila MezzomoiD, Albanin Aparecida Mielniczki-PereiraiD, Tanise Luisa SauseniD, Jorge MarinhoiD, Rogério Luís CansianiD
Abstract
The principle and the techniques applied in DNA extraction play a pivotal role in the obtention of a purified genetic material. The present study investigates the efficiency of eight protocols in the DNA extraction of Hypostomus commersoni, an essential component of South American freshwater ichthyofauna. The quality of samples was assessed through spectrophotometry, gel electrophoresis, and PCR-RAPD amplification. The efficiency of DNA extraction was influenced both by the method applied and the target-tissue of choice. Higher concentrations and yield of DNA were obtained from ocular tissue, with a positive spectrum of incubation in lysis buffer for up to 36 hours after sample collection, using fresh tissues and in the presence of a high concentration of Proteinase K (20 mg.ml-1). In these conditions, samples were successfully amplified. To date, there is no record of description for the parameters analyzed in this work, neither the description of RAPD markers for the species H. commersoni.
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