用聚乙二醇分离富集microRNA的方法探讨

目的探讨用聚乙二醇（polyethylene glycol，PEG）溶液分离富集miRNA的操作方法和分离富集效果，并与Ambion公司的miRNA分离试剂盒分离效果进行比较。方法用PEG溶液和Ambion公司的miRNA分离试剂盒从肝脏组织总RNA中分离富集miRNA，用变性琼脂糖和变性聚丙烯酰胺凝胶电泳鉴定分离效果，并在富集的miRNA中用RT—PCR扩增miR-122以鉴定是否有效地回收了miRNA。结果PEG和Ambion公司的miRNA分离试剂盒都能有效地富集miRNA，PEG富集的RNA片段比Ambion公司的试剂盒的大。结论PEG溶液能有效地分离富集miRNA，和Ambion公司的miRNA分离试剂盒分离效果相当，并具有操作简便、快捷及成本低廉的优点。

Isolation and Enrichment of MicroRNA by use of Polyethylene Glycol

Objective To develop approaches for isolating and enriching microRNA with Poly- ethylene Glycol （PEG）, in comparison with the microRNA isolation kit produced by Ambion com- pany. Methods microRNA isolation and enrichment were performed in liver tissue with PEG and Ambion microRNA isolation kit. The products produced with two methods were evaluated by electro- phoresis on denaturing agarose gel and denaturing polyacrylamide gel. miR-122 was detected by RT- PCR in the enrichment fraction to verify the recovery of microRNA in isolation process. Results PEG method was able to successfully enrich microRNA, similarly to Ambion microRNA isolation kit. The RNA fragment enriched by PEG was larger than that obtained by Ambion microRNA isola- tion kit. Conclusion PEG can enrich microRNA as efficiently as Ambion microRNA isolation kit, with advantage of easy and quick manipulation and low cost.