酚/氯仿法和盐析法提取人类外周血基因组DNA方法的比较

摘要

目的 比较两种不同方法提取的基因组DNA的纯度、产量和后续实验效果.方法 采用酚/氯仿法和盐析法两种方法直接从全血中提取基因组DNA,并采用电泳、PCR和基质支持的激光释放/电离飞行时间质谱分析(MALDI-TOF MS)方法进行检测.结果 酚/氯仿法提取的基因组DNA纯度高于盐析法(P＜0.001),但两组纯度平均值都高于1.80;没有发现两种方法提取的DNA浓度存在差异(P=0.819).两组DNA在电泳中都没有出现明显拖尾现象,均很容易扩增出胰岛素诱导基因2片段,并且在MALDI-TOF MS检测中,两组DNA样本结果没有明显差异.结论 酚/氯仿法和盐析法提取的基因组DNA质量无明显差异.相比酚/氯仿法,盐析法能简便、快速、无毒地进行DNA提取,适合于大规模的分子生物学实验.%Objective To detect the purity,quantity and follow-up experimental effects of DNA extracted with twornmethods. Methods Extract genomic DNA with phenol chloroform method and salting out method,and detect the DNArnsamples with electrophoresis, PCR and Matrix-Assisted Laser Desorption/ Ionization Time of Flight Mass Spectromerntry (MALDI-TOF MS). Results Purity of DNA extracted with phenol chloroform method was higher than that withrnsalting out method,with both mean purity higher than 1.80. The quantity was not significantly different between twornmethods (P=0. 819). The DNA samples did not smear on gel,and the PCR test of INSIG2 gene and MALD1-TOF MSrntest did not show difference between DNA isolated with two methods. Conclusion There was no difference betweenrnDNA samples isolated with phenol chloroform method and salting out method. Compared to phenol chloroform method,rnsalting out method was much simpler, more quick and harmless, and was applicable to molecular biology experimentsrnwith large scale.