An improved protocol suitable for polymorphism studies in Chinese jujube ( Ziziphus jujuba Mill.).

摘要

An efficient and stable AFLP protocol for Chinese jujube was established and validated with several cultivars. Major steps influencing in AFLP procedure were the extraction and purification of genomic DNA, restriction and ligation of adapter into the restricted genomic DNA, pre-amplification reaction, selective amplification reaction, electrophoresis and sliver-staining. The genomic DNA extracted by standardized modified CTAB method was suitable for downstream process in AFLP procedure. Four hundred ng DNA was digested with 3 units of EcoRI and MseI enzymes respectively and incubated at 37 degrees C for 5 h, the digested DNA fragment was diluted to 5 times and used as templates for pre-amplification, and the pre-amplification products were diluted to 10 times and used as templates for selective amplification, which was found suitable for Chinese jujube in AFLP. The electrophoresis and silver-staining were done as per normal procedure. This standardized method had also been validated with seven cultivars belonging to Chinese jujube.