【摘要】 在RNA干涉中,转染试剂的用量与siRNAs的量成正相关关系,但因转染试剂对细胞有一定的毒性作用,超过一定用量可以引起细胞形状改变、脱落、甚至死亡,直接影响着干涉的效率,所以确定转染试剂的优化用量,是RNA干涉试验中的重要环节.将不同用量的RNA iFectTM和SuperFect两种转染试剂分别加入鸡胚成纤维(CEF)细胞、293细胞、PK细胞和MDCK细胞中,在4h和24h后观察细胞的活力情况,比较了不同转染试剂对不同细胞的影响.结果表明:相同用量的两种转染试剂对293细胞、PK细胞和MDCK细胞的生长的影响远远小于对CEF细胞的影响;两种转染试剂用于外源小分子转染时,建议在293细胞、PK细胞和MDCK细胞上的用量不超过3u l;两种转染试剂对CEF细胞的毒性较大,不适合用于转染.更多还原

【Abstract】 In the RNA interference,the dosage of siRNAs has positive correlativity with that of transfection reagent.However,transfection reagents have a great influence on the growth of cells,including cells’ falling off,morphological change,even dying,etc.,when the dosage of the transfection reagent exceeds to some extend.So it is a very important process to RNA interference that ascertaining the optimization dosage of transfection reagent.In the test,the author added different dosages of two transfection reagents RNAiFect^TM and SuperFect to CEF cells、293 cells、PK cells and MDCK cells then observed the growth of these cells after 4h and 24h.The results showed that the growth of the CEF cells were greater effected than that of 293 cells、PK cells and MDCK cells when the same dosages of the two kinds of transfection reagents were used.It is recommended that when using the two transfection reagents to transfect siRNAs molecule into 293 cells、PK cells and MDCK cells,the dosage would be better under 3ul and they aren’t applicable transfection reagents for CEF cells,because of their great toxicity.更多还原