Comparación de protocolos de extracción de ADN con muestras de aleta y larva de peces: extracción modificada con cloruro de sodio

Abstract

The use of appropriate sampling methods, the type of tissue or sample and the use of viable DNA extraction protocols are critical issues in studies based on PCR. In an attempt to identify a simple, reproducible, inexpensive and non-toxic method for obtaining high quality and quantity genomic DNA from fi sh fi n and larvae samples, modifi ed salt extraction protocol and modifi ed phenol-chloroform extraction protocol were compared. The samples obtained from different fi sh species (Brycon orbignyanus, Piaractus mesopotamicus, Oreochromis niloticus, Leporinus elongatus and Prochilodus lineatus) using salt extraction showed good DNA quality and quantity (DNA/RNA relationship 1.8-2.2). These DNA samples were successfully used in the amplifi cation of RAPD (Random Amplifi ed Polymorphic DNA) and microsatellite molecular markers, demonstrating the same effectiveness of this protocol in comparison with modifi ed phenol-chloroform extraction protocol. This DNA extraction procedure constitutes an alternative and effi cient replacement for previous protocols for improving fi sh molecular studies.