%0 Journal Article
%A *
%A LIANG Yuan-Fa
%A LINGHU Chang-Di
%A LIU Guo-Min
%A TIANG Yong-Hui
%A YAN Dong-Hai
%A ZHENG Dao-Jun
%T Studies on Optimization of ISSR Amplified Conditions in Kudingcha Species in Oleaceae of China
%D 2009
%R 10.7525/j.issn.1673-5102.2009.02.019
%J Bulletin of Botanical Research
%P 234-241
%V 29
%N 2
%X The influential factors of ISSR for Kudingcha species in Oleaceae of China were systematically studied, a set of stable ISSR-PCR reaction parameters was established. 10 effective ISSR primers were selected out, and with them 21 test germplasm materials from 8 Kudingcha species in Oleaceae of China were examined for the repeatability and polymorphism of the optimized reaction conditions. The optimum ISSR-PCR reaction system was that 2.5 μL 10×PCR buffer, 2.0~3.0 mmol·L-1 MgCl2, 150~300 μmol·L-1 dNTPs, 1.0~1.5 Taq polymerase, 0.4~0.5 μmol·L-1 primers, and 5~320 ng DNA template were contained in 25 μL reaction solution. The optimized amplification program was that pre-denaturing at 94℃ for 4 min, then denaturing at 94℃ for 40 s, primer annealing at 50℃~54℃ for 45 s, extension at 72℃ for 120 s, for 35 cycles, at last extension at 72℃ for 8 min. The productions were stored at 4℃. The optimized ISSR-PCR reaction system was suitable for the study of genetic diversity and relationship of Kudingcha germplasm resources in Oleaceae of China.
%U https://bbr.nefu.edu.cn/EN/10.7525/j.issn.1673-5102.2009.02.019