Simplified and versatile method for isolation of high-quality RNA from pancreas.
Simplified and versatile method for isolation of high-quality RNA from pancreas.
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DOI：
10.2144/0000113862
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发表时间：
2012-05
期刊：
影响因子：
2.7
通讯作者：
Uc A
中科院分区：
文献类型：
--
作者：
Griffin M;Abu-El-Haija M;Abu-El-Haija M;Rokhlina T;Uc A
从胰腺中分离高质量的RNA是具有挑战性的，因为该器官含有大量的核糖核酸酶，并且在收获后立即进行自溶。本文介绍了一种简化的胰腺核糖核酸酶抑制剂灌流方法。这项技术始终从储存的胰腺样本中提取高质量的RNA，适合于分子生物学应用，包括定量实时RT-PCR。其产量与立即从胰腺中提取的RNA相当，但优于从快速冷冻或浸泡在核糖核酸酶抑制剂中的储存样品中提取的RNA。与原位导管灌注技术相比，我们的方法不会造成组织学伪影。
Isolation of high-quality RNA from pancreas is challenging because the organ contains large quantity of RNAses and it undergoes autolysis immediately upon harvest. A simplified perfusion method of pancreas with an RNAse inhibitor is described. The technique consistently yields high-quality RNA from stored pancreas samples that is suitable for molecular biology applications, including quantitative real-time RT-PCR. The yield is comparable to RNA isolated from the pancreas immediately, but superior to RNA isolated from stored samples that were snap frozen or immersed in an RNAse inhibitor. Compared to the in situ duct perfusion technique, our method does not cause histological artifacts.