摘要
目的通过对不同方法的比较建立一种遗传工程鼠基因组DNA鉴定提取最简单快速的方法。方法以大鼠和小鼠的鼠尾及脚趾为实验材料,采用酚氯仿、煮沸法、涡旋离心法提取基因组DNA;通过琼脂糖凝胶的方法检验DNA质量,通过对时间的统计分析比较3种方法提取DNA的速率。利用PCR技术检测DNA的扩增效果。结果煮沸法、涡旋离心法和酚氯仿提取的DNA比较,电泳条带均较弱。涡旋离心法所耗费的时间最短,煮沸法次之,酚氯仿法耗时最长。除了煮沸法外,涡旋离心法和酚氯仿方法提取的DNA在用于PCR检测时都能得到清晰的目的条带。结论在使用PCR进行基因型初步鉴定时,涡旋离心法是三种方法中最简单快速的首选方法。
Objective To find out a simple and rapid method for DNA extraction and detection in genetic engineering mouse and rat genome DNA by comprising different methods. Methods The tails and toes from rats and mice were as experimental material, genomic DNA were extracted by using phenol chloroform, boiling method and vortex centrifugation method. Concentration of DNA were measured by electrophoresis. The DNA quality was compared from the same size of tail and toe, the advantages and dis- advantages and maneuverability were evaluated among three methods, and effect of DNA amplification was detected by using PCR. Results The Concentration of DNA were lowest in vortex centrifugation method compared with that in the other 2 methods. Vortex centrifugation spent the shorter time than boiling method and phenol chloroform method was the longest one. Except the boiling meth- od. The DNA extracted by the Vortex eentrifugation method and the phenol chloroform method can obtain a clear target band for PCR detection. Conclusion vortex centrifugation was the most simple and fast method＇to preliminary appraisal the genotype identifica- tion by PCR.
出处
《解剖学研究》
CAS
2017年第6期479-482,共4页
Anatomy Research
基金
辽宁省科技计划项目(2015408002)
