%0 Journal Article
%A CHEN Da-Xia
%A LI Long-Yun
%A LU Cheng
%A ZHONG Guo-Yue
%A QU Xian-You
%A PENG Rui
%T Study on Optimization of ISSR Reaction Conditions for Coptis Chinensis Franch.
%D 2007
%R 10.7525/j.issn.1673-5102.2007.01.015
%J Bulletin of Botanical Research
%P 77-81
%V 27
%N 1
%X Based on the genomic DNA extracted from Coptis Chinensis Franch., this paper presented the effect of the main reaction system elements(Mg2+、dNTP、primer、template DNA、Taq DNA polymerase) and hot-cycle parameter(annealing temperature、cycles、denaturing time、annealing time and extension time) on ISSR-PCR which were tested by single or dual factor experiment, respectively to determine its optimal levels. A reaction system and amplified procedure suitable for Coptis Chinensis Franch were established,that is, 25μL amplification reactions system containing 1×PCRbuffer、1.5mmol·L-1 Mg2+、200μmol·L-1 dNTP、0.3μmol·L-1 primer、40 ng template DNA、1 U Taq DNA polymerase.The optimal amplified procedure was as follows: after a pre-denaturing of 30s at 94℃, 35 cycles were performed with denaturing of 30s at 94℃, annealing of 1min due to denaturing temperature of different primer, extension of 1.5min at 72℃, a final extension step of 7 min at 72℃ and hold at -4℃. This optimal system laid the standardization program of the identification of Coptis Chinensis Franch. and the efficient use of its germplasm resource.
%U https://bbr.nefu.edu.cn/EN/10.7525/j.issn.1673-5102.2007.01.015