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植物中微小RNA和其他小RNA的Northern印迹分析

Northern Blot Analysis of microRNAs and Other Small RNAs in Plants.

作者信息

De la Rosa Carlos, Reyes José Luis

机构信息

Departamento de Biología Molecular de Plantas, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Mexico.

出版信息

Methods Mol Biol. 2019;1932:121-129. doi: 10.1007/978-1-4939-9042-9_9.

Abstract

The study of regulatory small RNAs, such as siRNAs and microRNAs in plants, has necessitated methods tailored to their unique features. Their analysis demands the use of sensitive and quantitative methods for their detection. The use of Northern blot hybridization offers an attractive alternative to address qualitative as well as quantitative features. We highlight the advantages and shortcomings of this method and offer a detailed description of the techniques that best work in our hands, considering their use for the study of several small RNAs in multiple samples. We enumerate relevant details as well as cautionary comments in cases where we have detected potential difficulties.

摘要

对植物中诸如小干扰RNA(siRNA)和微小RNA(miRNA)等调控性小RNA的研究,需要有适合其独特特征的方法。对它们的分析需要使用灵敏且定量的检测方法。Northern印迹杂交的使用为解决定性和定量特征提供了一种有吸引力的替代方法。我们着重介绍了该方法的优缺点,并详细描述了在我们手中效果最佳的技术,同时考虑到它们在多个样本中用于研究几种小RNA的情况。我们列举了相关细节以及在检测到潜在困难的情况下的注意事项。

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