皱纹盘鲍SRAP反应体系的正交优化  

Optimization of SRAP-PCR System for disk abalone Haliotis discus hannai based on orthogonal design

作　　者：刘明泰 李杨[2] 刘小林[2] 许淑芬 常亚青[3] 

机构地区：[1]大连海宝渔业有限公司,辽宁大连116045 [2]西北农林科技大学动物科技学院,陕西杨凌712100 [3]大连海洋大学农业部北方海水增养殖重点实验室,辽宁大连116023

出　　处：《大连海洋大学学报》2013年第1期12-16,共5页Journal of Dalian Ocean University

基　　金：国家"863"计划项目(2006AA10A411)

摘　　要：采用L16(45)正交试验设计方法,对影响皱纹盘鲍Haliotis discus hannai SRAP反应体系的Mg2+浓度、dNTPs浓度、引物浓度、模板DNA以及Taq酶活性进行了优化,建立了适用于皱纹盘鲍的SRAP反应体系。优化后的反应体系(10μL)包括Mg2+1.5 mmol/L,dNTPs 0.15 mmol/L,引物0.2μmol/L,Taq酶活性0.25 U,模板DNA 20 ng。采用不同模板和引物对体系进行验证,表明优化后的反应体系能够高效地扩增出可识别条带,为进一步应用SRAP标记对皱纹盘鲍进行遗传分析奠定了基础。A L16（45） orthogonal test was employed in the present study to evaluate the influences of five important factors such as concentration of Mg2＋,dNTPs,primers,templates and activity of Taq DNA polymerase influencing amplification of SRAP-PCR,and optimization of SRAP-PCR System for disk abalone Haliotis discus hannai.The results showed that the optimum concentrations were found to be 1.5 mmol/L in Mg2＋,0.15 mmol/L in dNTPs,0.2 μmol/L in primer 0.25 U in activity of Taq enzyme,DNA polymerase and 20 ng in template DNA in a total volume of 10 μL reaction solution.The system was proved by other primers and template test to amplify reliable bands and to be used in the genetic analysis of disk abalone.

关 键 词：皱纹盘鲍 SRAP 正交试验设计 分子标记 反应体系 

分 类 号：Q953.1[生物学—动物学]