摘要
SSCP技术易受上样缓冲液、变性的时间及温度、电泳条件等诸多因素的影响。本研究对上样缓冲液、变性的时间及温度、电泳条件等影响因素进行了研究,结果显示:上样缓冲液中不添加甘油、用量为PCR产物体积的1.5～3倍、变性温度98℃、变性时间10～15min、电泳缓冲液为0.5×TBE,常温下恒功率80W(2.67W/cm)电泳2 h左右,单链DNA条带清晰,易于识别。
Single Strand Conformation Ploymorphism（SSCP） technique is easy to be affected by a good 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 wheat.The results were as follows when volume of the loading buffer without glycerol was 1.5～3 times of the volume of the PCR buffer;denaturalization temperature was 98℃;denaturalization time was 10～15 minutes;electrophoresis buffer was 0.5×TBE;the electrophoresis temperature was about 25℃;the electrophoresis power was 80W（2.67W/ cm）,the bands of the Single Strand DNA was clear and easy to read.
出处
《核农学报》
CAS
CSCD
北大核心
2007年第4期333-338,共6页
Journal of Nuclear Agricultural Sciences
基金
国家自然科学基金项目(30671302)
