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Using PCR-SSCP as tool to detect polymorphism in tapirs

Abstract:

Kocher et al.(1989), Goebel et al.(1999), Palumbi et al.(1991), Rueda & Morales (2005), Norton & Ashley (2004) and Richards et al.(1998). Following PCR, 8-10 μL of the samples were mixed with 4 μL of SSCP dye (formamide, bromofenol blue 0.25%, xilen-cianol 0.25% and sucrosa 40%), denatured at 95ºC for 5 minutes and cooled on ice for 5 minutes. Samples were run in 8, 10, and 12% acrylamide-bisacrylamide (49: 1) gels at 280-600V over 3-8 hours. Variants were recognized after Silver Nitrate staining as described by Sambrook et al (1989). The resolution of the gels has allowed us to resolve three control region variants, four 12S, one 16S, three cytB, and three COI, showing a high polymorphic variability among tapirs sampled. However preliminary our results, we are confident in the utility of the technique for tapir genetic studies. Studies in horses, for instance, have detected similar levels of polymorphism …