三种血液基因组DNA提取方法的比较研究

目的比较采用3种不同DNA提取方法提取豚鼠血液基因组DNA之间的差异。方法分别用新鲜抗凝血Na I手提法、血凝块手提法和试剂盒DNA提取法提取20只豚鼠血液基因组DNA,用分光光度计测定、琼脂糖凝胶电泳、PCR扩增等方法对提取的DNA片段的完整性、浓度、纯度和用于PCR扩增的效果等方面进行比较研究。结果新鲜抗凝血Na I手提法提取的DNA完整性、浓度和纯度均为最高;血凝块法提取的DNA浓度和抗凝血提取的DNA浓度接近但纯度最低,有一定程度的断裂和降解;试剂盒法提取的DNA浓度最低,纯度和完整性处于中间,但三种方法提取的DNA都可用于PCR扩增反应。结论 3种方法都可以提取基因组DNA并且所提取的DNA均能满足后续PCR扩增实验的要求,但每种方法各有利弊,可根据具体的条件选择适宜的方法。

Comparison of Three Methods for Blood Genomic DNA Extraction

Objective To compare the differences of three methods for DNA extraction from guinea pigs blood genomic DNA. Method Fresh anticoagulant NaI manual method, Blood clot manual method and DNA extraction kit method were used to extract blood genomic DNA from 20 guinea pigs. The integrity , concentration , purity and the effects of PCR amplification of obtained DNA were compared by Spectrophotometer,agarose gel electrophoresis, PCR amplification and other methods. Result The highest DNA integrity, concentration and purity could be achieved by employing fresh anticoagulant NaI manual method. The DNA concentration of blood clot manual method was approximate with the DNA concentration of fresh anticoagulant NaI manual method, but the purity was the lowest,and have some degree of breakage and degration. The DNA concentration obtained by DNA extraction kit was the lowest, integrity and purity were in the middle. There was no differences in PCR amplification results of the three methods. Conclusion The genomie DNA could be extracted by the three methods and could meet the requirements of subsequent PCR amplication, but each of them has its advantages and disadvantages, researchers could choose the suitable method according to specific conditions.