MicroRNA cloning from cells of the immune system.
MicroRNA cloning from cells of the immune system.
复制标题
从免疫系统细胞克隆 MicroRNA。
DOI：
10.1007/978-1-60761-811-9_5
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发表时间：
2010
期刊：
影响因子：
--
通讯作者：
Manjunath,N
中科院分区：
文献类型：
--
作者：
Wu,Haoquan;Neilson,Joel;Manjunath,N
MicroRNAs已经成为基因表达的重要转录后调节因子。小RNA克隆是发现新的microRNAs(MiRNAs)和分析miRNA表达的有效方法。此外，它揭示了在miRNA功能中可能重要的末端异质性。在这里，我们描述了一种优化的方案，可以从有限的起始材料中克隆小RNA。这通常是在高度纯化的免疫细胞或其他数量有限的主要细胞类型的群体中研究miRNAs的情况。用该方法克隆的小RNA具有miRNAs的典型特征--5‘-PO4和3’-OH基团，因此大多数克隆的小RNA都是miRNAs。
MicroRNAs have emerged as – important posttranscriptional regulators of gene expression. Small RNA cloning is a powerful method to identify new microRNAs (miRNAs) and to profile miRNA expression. In addition, it reveals end heterogeneity that may be important in miRNA function. Here, we describe a protocol that is optimized to clone small RNAs from limited amounts of starting material. This is often the case for studying miRNAs in a highly purified population of immune cells or other primary cell types with limited numbers. The small RNAs cloned with this protocol will have a 5′-PO4and 3′-OH group, typical features of miRNAs, so majority of the cloned small RNAs will be miRNAs.