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 markers 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.
Highlights
A crucial step for high-quality genetic analysis is considered to be a satisfactory DNA extraction
Our study has shown that a method that uses in-house produced reagents instead of commercial kits for DNA extraction can generate samples with sufficient quality to be used in molecular analysis, as PCR-RAPD amplifications
The relevance to describe an efficient method for DNA extraction for the species H. commersoni is justified by the fact that obtaining high-quality samples is crucial for studying organisms at the genetic level
Summary
A crucial step for high-quality genetic analysis is considered to be a satisfactory DNA extraction. This purpose is not always easy to achieve, since different biological compounds, such as lipids and proteins, can act as contaminants and interfere in the final quality of the product (Bauer and Patzelt, 2003). The principles and techniques applied in DNA extraction play a pivotal role in obtaining a considerable and purified yield of this molecule (Muhammad et al, 2016).
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