%0 Journal Article
%T Optimizing the Reaction System of DDRT-PCR by Positive-cross Test<br>正交法整体优化差异显示反应体系
%A LIU Shu-Fang~
%A <br>柳淑芳
%A 杜立新
%A 朱靖
%A 王爱华
%J 遗传
%D 2004
%I 
%X The method of mRNA Differential display PCR (DDRT-PCR) is used to identify differentially expressed genes widely. To improve further the efficiency and reproducibility of the method, the positive-cross test were performed to find the optional conditions of DDRT-PCR by analyzing the six critical parameters of dNTPs, MgCl 2, arbitrary primer, anchor primer, Taq enzyme, and template systematically. The experimental findings delineated the best possible DDRT-PCR conditions for a more reliable assessment of differential gene expression. Additionally, in this study, nondenaturing polyacrylamide gel electrophoresis (PAGE) and silver straining were adopted to reduce the false-positive ratio, and the reclaimed fragments were detected by reverse Northern analysis. Finally, we obtained the optimal condition for DDRT-PCR.
%K DDRT-PCR<br>正交优化
%K 非变性PAGE
%K 银染
%K 反向Northern
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=3E23F50E071DF18E21B2F5AEE4F1FA5E&aid=ACA6C9A28E351F73&yid=D0E58B75BFD8E51C&vid=96C778EE049EE47D&iid=B31275AF3241DB2D&sid=BC60A9A1D91963F5&eid=F7B726EE3ACCF328&journal_id=0253-9772&journal_name=遗传&referenced_num=5&reference_num=16