不同试剂盒提取质粒中内毒素水平的比较

Comparisons of Endotoxin Levels with Different Plasmid DNA Extraction Kits

目的比较几种不同质粒提取试剂盒提取质粒中内毒素水平。方法选用6种市售商业化试剂盒:QIAGEN质粒小提试剂盒Plasmid Mini Kit(12123),QIAGNE无内毒素质粒大提试剂盒EndoFree PlasmidMaxi Kit(12362),康为世纪高纯质粒中提试剂盒(CW0502),A厂家无内毒素质粒中提试剂盒,B厂家无内毒素质粒小提试剂盒,C厂家无内毒素质粒小提试剂盒(BSC01S2D)。分别提取质粒DNA,应用鲎试剂显色基质法和凝胶法检测所提质粒中内毒素水平。结果6种试剂盒提取质粒得率、A260/A280及A260/A230值均较好,但内毒素水平各不相同,其中各厂家标明无内毒素试剂盒提取质粒中内毒素水平相对较低,QIAGEN无内毒素质料大提试剂盒提取的质粒中内毒素小于0. 125 EU/μg。结论6种质粒提取试剂盒使用时应根据实际情况进行选择。

Objective To compare the endotoxin levels in plasmid purification with different plasmid DNA extraction kits.Methods Six commerical kits(QIAGEN Plasmid Mini Kit(12123),QIAGEN EndoFree Plasmid Maxi Kit(12362),CWBIO PurePlasmid Midi Kit(CW0502),EndoFree Plasmid Midi Kit,Endo-free Plasmid Mini Kit,Endo-free Plasmid DNA Mini Extraction Kit)were applied to purify plasmid,endotoxin levels were determined with the gel-clot technique and Limulus amebocyte lysate chromogenic method,respectively.Results The yields,A 260/A 280 and A/A 260 230 values of plasmid DNA extracted with the six kits were appropriate,the endotoxin levels were different,and the endotoxin levels extracted with endotoxin-free kits were relatively much lower.With QIAGEN EndoFree Plasmid Maxi Ki(t 12362),the endotoxin level was less than 0.125 EU/μg.Conclusion The endotoxin levels in plasmid DNA extraction with six kits were significantly different,which shall be selected according to the actual situation.