| RNA Isolation and Northern Blot Analysis
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Author:
Date:
2014-03-20
[Abstract] The northern blot is a technique used in molecular biology research to study gene expression by detection of RNA in a sample. With northern blotting it is possible to observe particular gene expression levels during differentiation, morphogenesis, as well as abnormal or diseased conditions. Here, we examine ATF3, ATF4, and GADD153 gene expression profiles by northern blot in Vero cells and H1299 cells after IBV infection. RNA was extracted in IBV (infectious bronchitis virus) infected cells and electrophoresis was used to separate the RNA sample. RNA was transferred from the electrophoresis gel to the blotting membrane by capillary transfer. Specific mRNA was detected with hybridization probes complementary to part of target sequence. The probes were prepared by RT-PCR and labeled by ...
[摘要] Northern印迹是在分子生物学研究中用于通过检测样品中的RNA来研究基因表达的技术。 使用Northern印迹，可以在分化，形态发生以及异常或疾病状况期间观察到特定的基因表达水平。 在这里，我们检查ATF3，ATF4和GADD153基因表达谱通过北部污点在Vero细胞和H1299细胞后IBV感染。 在IBV（感染性支气管炎病毒）感染的细胞中提取RNA，并使用电泳分离RNA样品。 通过毛细管转移将RNA从电泳凝胶转移到印迹膜上。 使用与靶序列的一部分互补的杂交探针检测特异性mRNA。 通过RT-PCR制备探针，并使用DIG标记试剂盒通过地高辛（DIG）标记。
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| Northern Blot of tRNA in Yeast
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Author:
Date:
2013-04-05
[Abstract] tRNAs are small RNAs around 70-90 nt. tRNAs are different from many other small RNAs in that they are very abundant, which makes it difficult to study their transcriptional regulation by traditional northern blot. Traditional Northern blot involves incorporation of radioactive nucleotides through polymerization, however, tRNA is too short for polymerization. Traditional Northern blot detects changes in RNA levels, however, tRNA are so abundant that small changes in their levels will escape detection. For these reasons, metabolic labeling by radioactive Uracil has been used instead. However, metabolic labeling can only examine changes in total tRNA, but cannot distinguish different types of tRNAs. The following protocol describes a method to examine individual tRNA gene transcription by ...
[摘要] tRNA是约70-90nt的小RNA。 tRNA与许多其他小RNA不同，因为它们是非常丰富的，这使得难以通过常规Northern印迹研究它们的转录调节。 传统的Northern印迹涉及通过聚合掺入放射性核苷酸，然而，tRNA对于聚合来说太短。 传统的Northern印迹检测RNA水平的变化，然而，tRNA是如此丰富，其水平的小变化将逃脱检测。 由于这些原因，已经使用了放射性尿嘧啶的代谢标记。 然而，代谢标记只能检查总tRNA的变化，但不能区分不同类型的tRNA。 以下方案描述了通过Northern印迹检查单个tRNA基因转录的方法。
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