Abstract:

Oncorhynchus mykiss is an important reared freshwater species, which has been cultured for more than thirty years in Gansu Province, China. Gansu golden trout is a new breed developed in Gansu, which had pass the National Aquaculture Species Identification Committee identifying and was recommend as the species reared in the northwestern areas. In this study, the DNA was extracted from the caudal fin of Gansu rainbow trout, and the conditions for AFLP analysis were optimized, including DNA concentration, dNTPs concentration, primer concentrati- on, denaturation time and annealing temperature.Then the AFLP reaction system was established. 100 ng genomic DNA was digested for 3 h at 37℃with34 U EcoR I and then 65℃for 5 h with 4 U Mse I , respectively; the product was ligated for 12 h with 1U ligase and was pre-amplificated; the products of pre-ampification was diluted 30 times, 8∶1 (w∶w) MseⅠ/ EcoRⅠselective primers for selective amplification; the amplification patterns were obtained clearly and stably. Finally, thirteen AFLP primer combinations that are suitable for Gansu Rainbow Trout cultivar were screened.