【摘要】 以冠突伪尾柱虫为材料 ,建立了一种适合于富含RNase和多糖的大型纤毛虫总RNA的提取方法。该方法采用异硫氰酸胍和 β 巯基乙醇联合快速变性 ,酚、氯仿和异戊醇抽提 ,同时在变性剂存在的条件下两次选择性地沉淀RNA ,从而有效地去除了DNA、蛋白质和多糖。所得RNA样品经电泳、紫外分光光度法检测和RT PCR分析 ,证实为完整、均一且无基因组污染的总RNA。这为构建冠突伪尾柱虫有小核系与无小核系之间的削减文库、筛选两细胞系差异表达的基因奠定了基础。更多还原

【Abstract】 The method for total RNA isolation from Pseudourostyla cristata was established using guanidinium isothiocyante, β-mercaptoethanol for denaturing, and water-sature phenol, chloroform and isoamyalcohole for extracting. The integrity and purification of the isolated total RNA were detected by using gel electrophoresis,RT-PCR and UV spectrophotometer. The quantity and quality of the total RNA extracted by this method is high enough for construct cDNA subtraction library and screen differential expressed genes.更多还原