摘要
目的检测SYBR-Gold前染色与后染色的优缺点。方法采用前染色、后染色的方法使用SYBR-Gold对双链DNA进行染色检测,EB染色作为参照,琼脂糖凝胶电泳为检测手段,凝胶成像系统拍照观察。结果SYBRGold其前染色与后染色对于双链DNA检测能力均强于EB染色,但是SYBR-Gold预混凝胶前染色其分辨效果及DNA迁移速度受到DNA浓度的影响;SYBR-Gold预混样品的前染色方式对于目的基因的判读误差较大。结论 SYBR-Gold不适合预混样品前染色,DNA样品浓度过大(大于250ng)不适合预混凝胶前染色,SYBR-Gold后染色是理想的染色方式。
Objective It is analyse two method for staining DNA fragments in agarosewith SYBR-Gold.Methods It used poststaining and prestaining to dye double strand DNA in agarose gel electrophoresis.The same way is used to stain DNA with EB as a control.And we analysed the results by the gel imaging system.Results We found SYBR-Gold was much more sensitive than EB to detect double strand DNA.But the effect of resolution and the mobility of DNA fragments were affiliated to the concentration of the samples in precasting SYBR-Gold in gels.And it was low accurate in measuring the size of DNA fragments through the agarose gel with adding SYBR-Gold in loading buffer.Conclusion We do not recommend that you stain the sample with SYBR-Gold before agarose gel electrophoresis.It is also not suitable to precast SYBR-Gold in gels when the concentration of samples is too high.However,the poststain with SYBR-Gold is an ideal method.
出处
《中国实验诊断学》
2015年第2期194-198,共5页
Chinese Journal of Laboratory Diagnosis
基金
国家自然科学基金项目(81100852)
