The Purity Identification Of Capsicum Hybrid With RAPD Markers

With the rapid development of Molecular Biology, RAPD(Random Amplified Polymorphic DNA)has been applied widely in Biological fields. This thesis focused on the application of RAPD to the Purity identification of capsicum hybrid seeds and researched into the optimal conditions of RAPD.It was analyzed that methods to extract DNA template and different ingredient had effects on RAPD. The results showed that the best reaction system is :DNA(25⁵0ng), MgCl₂ concentration (2. 0mmol/L), primer (15-45ng), dNTP (0. 2mmol/L),Taq Polyme-rase (1. 0¹. 5U) and the last reaction Procedure is that the reaction Pre — denatured DNA at 94℃ for 5 min, denatured DNA at 94℃ for40sec, annealed primers at 37℃ for40 sec, extended primers at 72℃ for 90 sec, re- extended reaction at 72℃ for 7min.The variety homogeneity was identified and the Parental Polymorphisms wereanalyzed. The Paper identified the homogeneity of the Parent seeds and the hybrid seeds and analyzed the Polymorphisms of the Parents before screening the Primers. The results were that there were no false seeds in Parent seeds by using the primer S61 and S1002.The Primers, which were suitable for the Purity identification of capsicum hybrid seeds, were screened and the fingerprints of RAPD were established among the Parents and their hybrid seeds in capsicum. 6 Primers were screened from 100: there were 2 Primers of partial female type(S10, S320), 2 Primers of Partial male type(S61, S74) , 2 Primers of complementary type(S1002, S301).Using the primer S1002 ,we can found that the Parental seed were all pure and there were 5 fault seeds (2, 7, 36, 62,65) in hybrid seeds.As a result, the Paper shows that RAPD can be applied to Purity identification of the capsicum seeds and that RAPD has good reliability and accuracy after the reaction system and the reaction Procedure are established.