Comparative evaluation on different methods to extract endophytic bacteria DNA from Diaphorina citri Kuwayama
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摘要: 通过分析细菌16SrDNA基因序列的PCR扩增与克隆测序结果,比较传统方法和试剂盒法对柑橘木虱Diaphorina citri Kuwayama内生菌DNA模板的提取效果。试验表明,4种方法中,FastDNA试剂盒方法提取的DNA模板较适合于后续柑橘木虱内生菌16SrDNA的扩增,且扩增产物可以很好地用于克隆测序;传统提取方法经济节约,但提取的DNA量及纯度较低;试剂盒提取方法,操作简单,质量稳定,易标准化,但成本较高。4种方法各具优缺点,用FastDNA试剂盒提取的DNA模板,其PCR产物能满足克隆需要。Abstract: To find a better method for the extraction of endophytic bacteria DNA from Asian Citrus Psyllid,Diaphorina citri Kuwayama,we compared the traditional methods and FastDNA Kits.The result showed that the DNA template extracted with the method of FastDNA Kits(MP Biomedical)was more suitable for PCR amplification,and the amplification product could be well used for cloning and sequencing.In the contrast,although the other three methods were cheaper,the amount and purity of DNA extracted with them were decreased.Our results suggested that the extraction with the FastDNA kits was stable and convenient for operation and standardization but expensiveness.In summary,four methods had respective advantages and disadvantages,and FastDNA kits provided more stable DNA for PCR and sequence cloning.
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Key words:
- Diaphorina citri Kuwayama /
- endophytic bacteria /
- total DNA /
- extraction
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