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Author:
Date:
2013-04-05
[Abstract] To examine gene expression, Northern blot or Real-Time PCR can be used to detect low abundant RNA such as mRNA. However, for high abundant RNAs such as rRNA and tRNA, Northern blot will not be able to discriminate the newly synthesized RNA from total RNA. Therefore, metabolic labeling is necessary to evaluate the expression of rRNA and tRNA genes. In this protocol, I describe a step-by-step method for labeling yeast RNA with radioactive uracil and examine the synthesis of these high abundant RNAs.
[摘要] 为了检查基因表达，可以使用Northern印迹或实时PCR来检测低丰度RNA如mRNA。 然而，对于高丰度的RNA如rRNA和tRNA，Northern印迹将不能区分新合成的RNA与总RNA。 因此，代谢标记是必要的，以评估rRNA和tRNA基因的表达。 在这个协议，我描述了一个分步的方法，用放射性尿嘧啶标记酵母RNA，并检查这些高丰度RNA的合成。
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