萃取和保存条件对人外周血细胞RNA提取效果的影响  

Effects of different conditions for extraction and storage on RNA extraction from peripheral blood cells

作　　者：吴苗[1] 闫林[1] 张红梅[1] 

机构地区：[1]山西医科大学公共卫生学院环境卫生教研室,山西太原030001

出　　处：《环境与健康杂志》2016年第3期249-251,共3页Journal of Environment and Health

基　　金：国家自然科学基金(81001285);太原钢铁集团卫生处资助项目(2014)

摘　　要：目的比较不同处理和保存条件下外周血RNA提取效果,找出最优的提取方案。方法外周全血去除血浆后,经氯仿萃取两次,加入不同体积比的RNAiso Plus试剂,对RNA进行提取。在此基础上,比较不同保存条件对RNA提取效果的影响,保存条件分别为:4℃保存24 h、72 h、10 d,-80℃保存10 d、30 d。通过紫外分光光度法测定RNA纯度和浓度,比较RNA提取效果,找出最优提取方案。结果与氯仿萃取一次相比,氯仿萃取两次提取的RNA纯度和浓度均升高,差异有统计学意义(P<0.05);血细胞与RNAiso plus试剂体积比为1∶1时提取效果最好;4℃保存10 d内的RNA提取效果优于其他保存条件。结论血细胞经氯仿萃取两次提取效果更好;加入等体积的RNAiso plus试剂提取效果最好;血细胞可在4℃稳定保存长达10 d,其RNA不被降解。Objective To find out the optimal RNA extraction protocol for peripheral blood cells by comparison for the effects of different treatments and storage conditions on RNA extraction. Methods The blood cells were obtained by removing plasma from peripheral whole blood with centrifugation, RNAiso Plus reagent was added to extract RNA, chloroform treatment was performed once and twice, and extration by different volumes of RNAiso Plus reagent were performed also. Afterwards, the influence of different storage conditions on RNA extraction were observed, which included 4 ℃ for 24 h, 4 ℃ for 72 h, 4 ℃ for 10 d,-80 ℃ for 10 d,-80 ℃ for 30 d. RNA＇s extraction effect were judged based on the purity and concentration, which were measured by the ultraviolet spectrophotometry. Results The purity and concentration of RNA were significantly increased for the twice chloroform treatment compared with the once treatment, and were the highest in 1 ∶1 volume ratio of blood cells to RNAiso Plus reagent. The purity and integrity of RNA were well maintained in 4 ℃ up to 10 days compared with those in-80 ℃.Conclusion For peripheral blood cells RNA extraction, it should be better to conduct twice chloroform treatment and the equal volume of RNAiso Plus, and the integrity of RNA inside blood cells can be maintained up to 10 days in 4 ℃.

关 键 词：外周血细胞 RNA提取 纯度 浓度 比较 

分 类 号：O657.3[理学—分析化学]