摘要
目的 建立外周血RNA的提取方法 ,为用逆转录 -聚合酶链反应检测乳腺癌外周血中微转移打好基础。方法 从 30例乳腺癌患者的冰冻保存和新鲜全血标本 ,用两种不同方法分离外周血有核细胞 ,提取RNA ,测定RNA产量及RNA的完整性。结果 从冰冻保存和新鲜全血标本 ,提取的总RNA的OD值有显著性差异。从新鲜全血中提取RNA的产量高于从冰冻保存的全血中提取的RNA的产量。但是 ,电泳时均有 2 8S与 1 8S的条带。结论 用逆转录 -聚合酶链反应时 ,以从新鲜全血中提取RNA为宜 ,但为留取标本的方便 ,也可用冰冻保存的全血标本 。
Objective To establish methods of extracting total RNA of nucleated cells in peripheral blood for detecting breast cancer micrometastases with reverse transcription polymerase chain reaction (RT-PCR).Methods Peripheral blood from 30 breast cancer patients were gone two chares.One of them was freezed and seperated nucleated cells,and another was seperated nucleated cells directly Two groups were extracted total RNA and estimated their qualities.Results There was difference between the two shares of RNA OD values.The RNA qualities were more from non-freezed than from freezed peripheral blood.But after they were run 1% denaturing agarose-formalhyde gels electrophoresis,all of them were intact 28S and 18S RNA of total RNA were clearly seen.Conclusion It is better that extracted total RNA from non-freezed peripheral blood.But the two methods can be carreied out extraction total RNA for RT-PCR.
出处
《兰州医学院学报》
2002年第4期9-10,共2页
Journal of Lanzhou Medical College
