霍乱弧菌基因组DNA提取方法的比较研究

Comparison of different methods to extract Vibrio cholerae Genomic DNA

目的目前细菌DNA提取方法众多,但由于提取原理和步骤的差别,获得DNA的浓度及纯度各不相同,本文通过霍乱弧菌基因组DNA提取方法的比较研究,为不同需求提供不同的DNA提取方法建议。方法本文采用4种不同DNA提取方法(酚-氯仿抽提法、CTAB法、试剂盒法和液氮研磨法)提取霍乱弧菌基因组DNA,并比较各种方法提取DNA的效果。结果酚-氯仿抽提法和CTAB法提取的基因组DNA效果相对较差,但成本低;液氮研磨法提取的基因组DNA效果大大改进;试剂盒法不仅操作简便,而且提取的基因组DNA效果最好。结论4种方法均适合霍乱弧菌的聚合酶链反应(PCR)检测,试剂盒法还适用于分子分型和基因组测序分析。

Objective DNA molecular diagnosis represents the development trend of the Vibrio cholerae detection.Effective extraction of the genomic DNA is a key step in the detection of V.cholerae by molecular biological methods.At present,various bacterial DNA extraction methods are used,however,the concentration and purity of DNA varied due to the different principle and procedure.Methods In this study,four different DNA extraction methods（phenol-chloroform extraction,CTAB,commercial kit test and liquid nitrogen grinding） were used to extract V.cholerae genomic DNA and the results were compared.Results The results showed that the efficiencies of phenol-chloroform extraction and CTAB were not high,but their costs were low.Compared with liquid nitrogen grinding by which the efficiency was improved,the commercial kit test still had the advantage of higher efficiency besides the easy operability.Conclusion Phenol-chloroform extraction,CTAB and liquid nitrogen grinding are only suitable for the PCR detection of V.cholerae,but the commercial kit test is suitable for the PCR detection,molecular typing and genomic sequencing of V.cholerae.