少量外周血提取RNA方法的研究

来源 :中国热带医学 | 被引量 : 0次 | 上传用户:infoerp2009
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目的建立从少量外周血中提取RNA的方法,为相关分子生物学及遗传学研究提供条件。方法从13例健康成人各抽取外周血3份,每份0.3ml,一份立即提取RNA,一份放置室温3d,另一份放置-20℃保存21d,提取RNA,测定RNA产量、RNA的完整性及用RT-PCR方法检测其对不同基因的表达。结果从新鲜全血中提取RNA的产量相对较高,与从室温及冰冻保存的全血中提取的RNA的产量相比有统计学意义。三组提取RNA均有良好的完整性,均可用于RT-PCR。结论在用少量外周血标本做RT-PCR等研究时,可以用PreAnlytix方法提取少量血中RNA,为留取标本的方便,也可用室温或冰冻保存的全血标本。 Objective To establish a method for RNA extraction from a small amount of peripheral blood and provide conditions for related molecular biology and genetics research. Methods Peripheral blood samples were collected from 13 healthy adults at 0.3ml each. One sample was immediately extracted from RNA, one was stored at room temperature for 3 days and the other was stored at -20 ℃ for 21 days. RNA was extracted and the RNA production and RNA integrity The expression of different genes was detected by RT-PCR. Results The yield of RNA extracted from fresh whole blood was relatively high, which was statistically significant compared to the yield of RNA extracted from room temperature and cryo-preserved whole blood. Three groups of RNA extraction have good integrity, can be used for RT-PCR. Conclusion In a small amount of peripheral blood samples for RT-PCR and other studies, PreAnlytix method can be used to extract a small amount of blood RNA for the convenience of taking samples can also be stored at room temperature or frozen whole blood samples.
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