一种使用夹板连接法直接检测微小RNA和其他小RNA的快速定量检测方法。
A rapid, quantitative assay for direct detection of microRNAs and other small RNAs using splinted ligation.
作者信息
Maroney Patricia A, Chamnongpol Sangpen, Souret Frédéric, Nilsen Timothy W
机构信息
Center for RNA Molecular Biology and Department of Biochemistry, Case Western Reserve University, School of Medicine, Cleveland, Ohio 44106-4973, USA.
出版信息
RNA. 2007 Jun;13(6):930-6. doi: 10.1261/rna.518107. Epub 2007 Apr 24.
Abstract
The discovery and characterization of microRNAs (miRNAs) and other families of short RNAs has led to a rapid expansion of research directed at elucidating their expression patterns and regulatory functions. Here, we describe a convenient, sensitive, and straightforward method to detect and quantitate specific miRNA levels in unfractionated total RNA samples. The method, based on splinted ligation, does not require specialized equipment or any amplification step, and is significantly faster and more sensitive than Northern blotting. We demonstrate that the method can be used to detect various classes of small regulatory RNAs from different organisms.
摘要
微小RNA（miRNA）及其他短RNA家族的发现与特性描述，促使针对阐明其表达模式和调控功能的研究迅速扩展。在此，我们描述了一种简便、灵敏且直接的方法，用于检测和定量未分级总RNA样本中特定miRNA的水平。该方法基于夹板连接，无需专门设备或任何扩增步骤，且比Northern印迹法显著更快、更灵敏。我们证明该方法可用于检测来自不同生物体的各类小调控RNA。
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2
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Biotechniques. 2006 Oct;41(4):420, 422, 424. doi: 10.2144/000112270.
3
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