三种人全血基因组DNA提取方法的比较

目的：比较改良酚一氯仿抽提法、盐析法、试剂盒法从人全血中提取基因组DNA的效果，以期建立一种快速、经济的提取高质量基因组DNA的方法。方法：分别用上述三种方法从人全血中提取基因组DNA，通过紫外分光光度计、琼脂糖凝胶电泳、聚合酶链式反应（PCR）、限制性内切酶酶切检测提取的基因组DNA的产量、纯度和质量。结果：改良酚一氯仿抽提法与试剂盒法提取的基因组DNA相比，DNA的产量有统计学差异，DNA的纯度无统计学差异，但试剂盒法提取的基因组DNA有较明显的降解现象：盐析法与改良酚．氯仿抽提法、试剂盒法相比，基因组DNA的产量和纯度都存在统计学差异，并且基因组DNA聚合酶链式反应（PCR）扩增的稳定性也明显劣于另外两种方法；三种方法提取基因组DNA均能进行限制性内切核酸消化。结论：改良酚一氯仿抽据取法是一种经济、快速、高效、稳定提取人全血基因组DNA的方法，适用于批量临床标本处理。

Comparing the Three Methods to Extract Human Genomic DNA from Whole Blood

Objective： To explore a rapid, economical method for extracting high quality genomic DNA expectably, the differences of improved phenol-chloroform method, salt-extraction method and kit method on extracting human genomic DNA fi＇om whole blood were investigated. Methods： Human genomic DNA was extracted by these three methods, respectively. The genomic DNA yield, purity and quality were measured by ultraviolet photometer, agarose gel electrophoresis, PCR amplification and digesting of restrictive endonu- clease. Results： The genomic DNA yield of improved phenol-chloroform method, salt-extraction method and kit method were 125.36± 9.39 ng.μL1, 97.26± 19.89 ng.μL-1, 383.14± 23.01 ng.μL-1, respectively. However, genomic DNA was not integrity with the kit method, and there was no significant difference in DNA purity between the improved phenol-chloroform method and the kit method. The salt-ex- traction method delivered significantly lower yields of DNA and DNA purity compared to other two methods. Moreover, PCR amplifica- tion was not stable with genomic DNA extracted by using the salt-extraction method. The genomic DNA extracted with these three meth- ods were all identified by digesting of restrictive endonuclease effectively. Conclusion： Improved phenol-chloroform method is an economi- cal, rapid, efficient and stable method for extracting genomic DNA fi＇om whole blood, and is applicable for large numbers of clinical sam- pies.