Optimization of RAPD-PCR Reaction System for Glycyrrhiza uralensis Based on Orthogonal Design  

甘草RAPD-PCR反应体系正交优化研究(英文)

作　　者：张增福 董建力[2] 李明[3] 

机构地区：[1]宁夏农业技术推广总站,宁夏银川750001 [2]宁夏农林科学院农业生物技术研究中心,宁夏银川750002 [3]宁夏农林科学院沙漠化治理研究所,宁夏银川750002

出　　处：《Agricultural Science & Technology》2011年第10期1427-1429,共3页农业科学与技术（英文版）

基　　金：Supported by Key Projects in the National Science&Technology Pillar Program in the Eleventh Five-year Plan Period(2006BAI06A15-11)~~

摘　　要：[Objective] The aim was to obtain the optimum RAPD-PCR reaction system for Glycyrrhiza uralensis.[Method] Orthogonal design was adopted to screen the suitable concentration of four major factors（dNTPs,primers,Taq polymerase and DNA template） in PCR reaction system.[Result] The optimal reaction system obtained by orthogonal design was 25 μl in total volume,containing 2.5 μl of 10×PCR buffer solution（include MgCl2）,2.5 μl of 10 mmol/L dNTPs,2 μl（100 ng） of DNA template,2 μl of 10 μmol/L primers,0.4 μl（5 U） of Taq polymerase;the optimum annealing temperature was 34 ℃.[Conclusion] Orthogonal design was an effective method for the optimization of RAPD-PCR reaction system for G.uralensis.[目的]建立一套适合甘草分子学研究的RAPD-PCR反应体系。[方法]以甘草种质为试材,采用正交试验法设计,对影响RAPD-PCR扩增的主要因素dNTPs、引物、Taq酶和DNA模板进行优化筛选。[结果]总体积25μl的甘草RAPD-PCR最佳反应体系为:10×PCR缓冲液(含MgCl2)2.5μl,10mmol/LdNTPs2.5μl,50ng DNA2μl,10μmol/L引物2μl,5UTaq酶0.4μl。对引物的退火温度进行了梯度筛选,34℃时扩增效果较好。[结论]进行甘草RAPD-PCR反应体系的正交优化非常有效。

关 键 词：Glycyrrhiza uralensis DNA RAPD-PCR Reaction system OPTIMIZATION Orthogonal design 

分 类 号：S567.71[农业科学—中草药栽培]