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

Abstract

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.

Cross-amplification of heterologous microsatellite markers in Piracanjuba

“…DNA was extracted following methodology by LOPERA-BARRERO et al (2008). Total DNA concentration was assessed by measuring samples by spectrophotometry PICODROP ® (Picodrop Limited, Hinxton, UK).…”

Genetic characteristics of Tambaqui broodstocks in the state of Rondônia, Brazil: implications on production and conservation

“…Several studies have revealed that the formation of broodstocks with low genetic variability may cause difficulties related to endogamy cycles, adaptation capacity and survival of offspring (LOPERA-BARRERO et al, 2010a;RODRIGUEZ-RODRIGUEZ et al, 2010;LOPERA-BARRERO et al, 2014) and may affect the genetic potential of the species (MOREIRA et al, 2007). Low genetic diversity is negatively correlated with adaptation potential to changes in environmental conditions (RASHID et al, 2012;LYNCH et al, 1995;LOEW, 2000).…”

“…The characterization of genetic variability is relevant for production management , establishment and maintenance of improvement (OLIVEIRA et al, 2012) and conservation programs (LOPERA-BARRERO, 2009) as an integral part in the formation, maintenance and correct use of broodstocks. Several studies have revealed that the formation of broodstocks with low genetic variability may cause difficulties related to endogamy cycles, adaptation capacity and survival of offspring (LOPERA-BARRERO et al, 2010a;RODRIGUEZ-RODRIGUEZ et al, 2010;LOPERA-BARRERO et al, 2014) and may affect the genetic potential of the species (MOREIRA et al, 2007).…”

Diversidad genética de piracanjuba usada en programas de repoblación con marcadores microsatélites

“…Para la extracción de ADN, se utilizó la metodología descrita por Lopera Barrero et al (2008a) Tris-Borato, 2 mmol L -1 EDTA) a 350 V (250 mA), durante 6 horas. Para visualizar los alelos microsatélites, se utilizó la coloración con nitrato de plata descrita por Bassam et al (1991), con algunas modifi caciones.…”

“…Según Povh et al (2008a), las introducciones de peces de forma no planeada pueden provocar una reducción de la variabilidad genética, por manejos genéticos y reproductivos inadecuados, y pueden provocar erosión genética en las poblaciones naturales receptoras de alevinos liberados. Así, con el propósito de implementar un programa de conservación efi ciente, se debe considerar la preservación genética de las poblaciones de peces nativos, para lo cual es importante obtener una identifi cación y caracterización de procedimientos genéticos y reproductivos utilizados en lotes de esta especie, cuando orientados a la repoblación (Sirol & Britto, 2006).…”