%0 Journal Article
%A BAO Tian-Zhong
%A GONG Ai-Qi
%A HU Yan-Ping
%A LI Rui-Xin
%A LI Yi*
%A PENG Bing-Yu
%A WANG Huan-Qiang
%A WANG Li
%T Establishment and Optimization of ISSR-PCR System for Hybrid Rapeseed
%D 2010
%R 10.7525/j.issn.1673-5102.2010.05.010
%J Bulletin of Botanical Research
%P 576-581
%V 30
%N 5
%X An orthogonal design was used to optimize the ISSR-PCR system for hybrid rapeseed at three levels of four factors (Mg<sup>2+</sup>, dNTP, primer and <em>Taq</em> DNA polymerase). Then, based on the optimal ISSRPCR amplification system, the concentration of DNA template and annealing temperature were selected. As a result, the optimal PCR (20 &mu;L) mixture contained 1.50 mmol&middot;L-1 Mg<sup>2+</sup>, 0.125 mmol&middot;L<sup>-1</sup> dNTP, 2.00 &mu;mol&middot;L<sup>-1</sup> primer, 0.50 U <em>Taq</em> DNA polymerase, 2.5 &mu;L 10&times;buffer and 40ng template DNA. The suitable annealing temperature of primer UBC891 was 54.2℃. With the optimal system of ISSR-PCR reaction, clear and steady bands were obtained in different individuals of Qingza No.3 and its male parent. The establishment of ISSR-PCR system could favor the studies on the purity and genuineness of hybrid rapeseed varieties by using molecular marker techniques.
%U https://bbr.nefu.edu.cn/EN/10.7525/j.issn.1673-5102.2010.05.010