%0 Journal Article
%A CHEN Jia-Li
%A CHEN Xuan
%A SUN Xiu-Xiu
%A YANG Li-Rong
%A YUN Yong
%A ZHENG Dao-Jun
%T Optimization of SCoT-PCR System and Screening of Effective Primers for <i>Castanopsis wenchangensis</i>,an Extremely Small Population Endemic to Hainan Province
%D 2020
%R 10.7525/j.issn.1673-5102.2020.01.010
%J Bulletin of Botanical Research
%P 66-72
%V 40
%N 1
%X The effects of DNA, primers and dNTPs, Taq DNA polymerase on the SCoT-PCR amplification results of <i>Castanopsis wenchangensis</i> were analyzed by single factor test and orthogonal test, and the SCoT-PCR system of <i>C.wenchangensis</i> was optimized. The polymorphism primer combination was screened based on the established reaction system, which provided the conditions for the study of SCoT molecular markers in the study of <i>C.wenchangensis</i> germplasm resources. The results showed that the influence of each factor on SCoT-PCR amplification was DNA > dNTPs > primer > Taq enzyme, and the optimum reaction system was as follows:when the total system was 20 μL, DNA content was 2.5 ng, primer concentration was 0.8 μmol&#183;L<sup>-1</sup>, dNTPs concentration was 0.2 mmol&#183;L<sup>-1</sup>, Taq content was 1 U, and the amplified products were clear and stable. In addition, 15 primers with good polymorphism and suitable for <i>C.wenchangensis</i> were screened out from 80 SCoT primers by this system, which provided theoretical basis and technical support for the related research of <i>C.wenchangensis</i> and other plants of Fagaceae by SCoT molecular marker technique in the future.
%U https://bbr.nefu.edu.cn/EN/10.7525/j.issn.1673-5102.2020.01.010