猪粪和土壤样品中微生物DNA提取方法的比较

猪粪施于土壤可能会对土壤微生物多样性造成影响,为选用同一种DNA提取方法用于土壤和猪粪微生物DNA的提取,该文采用了化学裂解法和试剂盒法同时从土壤和猪粪样品中提取微生物DNA,并对这两种方法的提取DNA的效果进行了比较。结果表明,试剂盒法不能用于提取土壤中的微生物DNA;可以从猪粪中提取到DNA,PCR扩增能得到目的产物,但重复性不高。化学裂解法提取的土壤微生物DNA浓度高但纯度低,纯化后纯度增加,但DNA有所损失,用于PCR扩增时结果不理想;处理猪粪样品,提取的DNA浓度较低但纯度较高,PCR扩增结果比较理想。由此可见,化学裂解法用来提取猪粪样品中的微生物DNA是可行的,但需寻求更好的土壤样品微生物DNA的提取方法。

Comparison of methods for microbial DNA extraction from swine feces and soil

The growth of the livestock industry resulted in the increase of feces and sullage,which caused consider- able environment pollution.Swine feces would affect the soil microorganism when it entered the soil.To study the influence of swine feces on soil microorganisms,the same method for microbial DNA extraction from soil and swine feces was the basis.The efficiency of two methods as chemical lysis method and DNA Isolation Kit method for microbial DNA extraction was compared in this experiment.The results show that microbial DNA from swine feces can be extracted by two methods.But microbial DNA of soil can't be extracted by the DNA Isolation Kit method.Microbial DNA extracted from swine feces can be used for PCR amplification,but the amplification result is not steady.Microbial DNA from soil is high in yield and low in purity when the chemical lysis method is used. By contraries,microbial DNA from swine feces is low in yield and high in purity.It shows that microbial DNA extracted from swine feces is more suitable for PCR amplification than from soil.After purification the extracted DNA samples from swine feces can be used for PCR amplification,using primers of the bacterial 16SrDNA gene, but not be the extracted DNA samples from soil.In conclusion,the chemical lysis method for microbial DNA ex- tracted from swine feces is feasible.But the better method for microbial DNA extracted from soil is needed to be studied.