赤眼蜂SSCP分子标记体系的优化  

Optimization of SSCP Technique in Trichogramma spp.

作　　者：董辉[1] 柳晓利[1] 钱海涛[1] 丛斌[1] 

机构地区：[1]沈阳农业大学植物保护学院,沈阳110161

出　　处：《沈阳农业大学学报》2011年第4期428-432,共5页Journal of Shenyang Agricultural University

基　　金：国家自然科学基金项目(30871674;30971962);"十一五"国家科技支撑计划项目(2006BAD0806-01);辽宁省高校重点实验室支持计划项目(2008S203);辽宁省教育厅科研项目(L2010488);沈阳农业大学教师科研基金项目(20091012)

摘　　要：以松毛虫赤眼蜂为试验材料,对SSCP分子标记体系的电泳条件、温度、变性时间、上样缓冲液等诸多影响因素进行了研究。结果表明：4℃恒温,7W恒功率下,上样缓冲液中不添加甘油、用量为PCR产物体积的0.5~3.5倍,变性温度95℃或98℃,变性时间10~15min,电泳缓冲液为1×TBE,电泳16h,为SSCP分子标记体系最佳的条件组合。Single strand conformation ploymorphism（SSCP）technique is easy to be affected by many factors such as loading buffer,denaturalization temperature and time,and electrophoresis factors.The influences of loading buffer,denaturalization temperature and time and electrophoresis factors（electrophoresis temperature,electrophoresis buffer and electrophoresis power）on SSCP technique were studied in Trichogramma spp,The results showed when volume of the loading buffer without glycerol was 0.5~3.5 times of the volume of the PCR buffer;denaturalization temperature was 95℃ or 98℃;denaturalization time was 10~15 minutes;electrophoresis buffer was 1 × TBE;the electrophoresis temperature was about 4℃;the electrophoresis power was 7W;the electrophoresis time was 16h;the bands of the single strand DNA were clear and easy to read.

关 键 词：赤眼蜂 SSCP 反应体系 

分 类 号：S476.3[农业科学—农业昆虫与害虫防治]