| Biotinylated Micro-RNA Pull Down Assay for Identifying miRNA Targets
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Author:
Date:
2017-05-05
[Abstract] microRNA (miRNA) directly associates with its target transcripts (mRNA). This protocol describes a method for detection of direct interaction between miRNA and mRNA. The result of interaction helps screening the specific target mRNAs for a miRNA.
[摘要] microRNA（miRNA）与其目标转录物（mRNA）直接相关。 该方案描述了检测miRNA和mRNA之间直接相互作用的方法。 相互作用的结果有助于筛选miRNA的特异性靶mRNA。
背景 MiRNA是小的非编码调控RNA。 MiRNA通常通过与靶mRNA中的互补序列结合来控制基因表达。 许多生物信息学和计算程序可用于预测miRNA靶标。 但是鉴定miRNA与靶mRNA的直接相关性的实验方法非常有限。 目前的方案可以非常有助于识别miRNA靶标（Phatak等人，2016）。
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| Determination of the Secondary Structure of an RNA fragment in Solution: Selective 2`-Hydroxyl Acylation Analyzed by Primer Extension Assay (SHAPE)
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Author:
Date:
2015-01-20
[Abstract] This protocol describes the methodology for the determination of the secondary structure of an RNA fragment in solution using Selective 2´-Hydroxyl Acylation analyzed by Primer Extension, abbreviation SHAPE. It consists in the very fast chemical modification of flexible and therefore possibly single-stranded nucleotides in a sequence-independent manner using benzoyl cyanide (BzCN), forming 2´-O-adducts. The modifications in the RNA are then analyzed by primer extension. Reverse transcriptase is blocked by the 2´-O-adducts formed. The advantage of the method is, first, that not each RNA ...
[摘要] 该方案描述了使用通过引物延伸（缩写为SHAPE）分析的选择性2'-羟基酰化来确定溶液中RNA片段的二级结构的方法。 其包括使用苯甲酰氰（BzCN）以序列非依赖性方式非常快速地化学修饰柔性且因此可能的单链核苷酸，形成2'-O-加合物。 然后通过引物延伸分析RNA中的修饰。 逆转录酶被形成的2'-O-加合物阻断。 该方法的优点是，首先，不是每个RNA分子研究，而是在延伸反应中使用的引物被标记，其次，在测序凝胶中分析得到的cDNA比修饰的RNA更稳定。
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| Protein-RNA ELISA Assay
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Author:
Date:
2012-09-05
[Abstract] Protein-RNA ELISA assay is an effective and quantitative method to study protein-RNA interactions in vitro. In this protocol we used recombinant 6x HIS tagged protein, but it works as well for non tagged proteins.
[摘要] 该实验方案的中文版正在准备中．．．
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