目的:发展一种简单快速经济的回收琼脂糖凝胶中DNA的方法。方法:将切的琼脂糖凝胶胶块放入嵌套Eppendorf管中捣碎,加入50μL机油,室温下12 000r/min离心5min,取大Eppendorf管中收集的液体跑胶检验,凝胶上包括代表100bp~2000bp不同大小DNA分子回收率的分子量标准DL2000。结果:DNA回收率可由加机油前的约35%提高为加机油后的45%-90%,回收效率的波动主要取决于DNA片段的大小、切下的含有DNA的胶块的大小及操作者的熟练程度。结论:该方法快速、简便、经济,具有良好的重复性与特异性,比许多国产的试剂盒更可靠。
Objective:To develop a simple,easy and inexpensive method for the rapid DNA recovery from agarose gel slice by using motor oil.Method: A 0.5ml Eppendorf tube was punctured in the bottom with a syringe needle for the opening of a hole.Then it was loosely packed with cotton wool,which was subsequently placed inside a capless 1.5 ml Eppendorf tube.Pieces of the gel slice were placed inside the small tube and mashed with pipet tips.Then it was added with motor oil,and subjected to a 5min centrifugation at 12000r/min at room temperature.Flowthrough was visualized on agarose gel.Result: DNA recovery rate could reach 45%-90%,showing a marked improvement over our previous method.Conclusion: This method is simple,rapid,economical,efficient,and more reliable than domestic DNA recovery kits.