Construction of AFLP reaction system in leaves of Castanea mollissima in the Yanshan Mountains

High quality DNA required for construction of AFLP analysis system was isolated with an improved method of CTAB from young leaves of Castanea mollissima. A silver-staining AFLP reaction system was constructed for genetic analysis of C. mollissima by optimizing DNA restriction-ligase reaction, pre-amplification and selective amplification.Resultsshowed that the system of restriction of DNA was better than restriction and link of DNA. The polymorphism bands were obtained from three primer pairs (E-AAC+M-CAA, E-AAC+M-CAT, E-AGT+M-CAT), but E-AGT+M-CAT could amplify more uniform and high sharp bands. Our study showed that the silver-staining AFLP was available for genetic diversification, variety identification, construction of molecular map, identification and classification.