【摘要】 目的探讨两种不同方法提取结核分枝杆菌总RNA,并在实验中对其进行优化。方法收集结核分枝杆菌培养物,分别用甲醇和玻璃粉裂解其细胞壁,然后加入Trizol提取结核分枝杆菌总RNA,用琼脂糖凝胶电泳检测其完整性,用Nano-drop2000检测其得率和纯度。结果玻璃粉裂解细胞壁法和甲醇裂解细胞壁法提取的结核分枝杆菌总RNA的获得率分别为6.124±1.144和13.437±1.767(P<0.01),纯度A260/A280的值分别为1.924±0.039和1.899±0.072(P>0.01)。结论玻璃粉裂解细胞壁法和甲醇裂解细胞壁法提取的结核分枝杆菌总RNA均未发生明显降解,其完整性均能满足后续实验的需要。用甲醇裂解法提取的结核分枝杆菌总RNA的获得率明显高于玻璃粉法。更多还原

【Abstract】 Objective To compare two methods(glass beads and methanol) for extracting the total RNA of Mycobacterium tuberculosis.Methods After being inactivated,the cultures of Mycobacterium tuberculosis were lysed with glass beads and methanol,and then the total RNA was extracted with Trizol.The integrity of the RNA was checked by electrophoresis through agarose gel and the concentration of RNA was measured spectrophotometerically.Results The quantities of the total RNA of Mycobacterium tuberculosis with glass beads and methanol were 6.124±1.144 and 13.437±1.767(P<0.01),respectively,and A260/A280 values were 1.924±0.039 and 1.899±0.072(P>0.01),respectively.Conclusion The total RNA of Mycobacterium tuberculosis with two methods can satisfy the demands in quality for microarry.The quantities of the total RNA extracted with methanol is higher than that with glass beads.更多还原