病毒的凝胶印迹杂交。

Gel Blot Hybridization for Viroids.

机构信息

Instituto de Biología Molecular y Celular de Plantas (UPV-CSIC), Consejo Superior de Investigaciones Científicas-Universitat Politecnica de Valencia, Valencia, Spain.

出版信息

Methods Mol Biol. 2022;2316:97-109. doi: 10.1007/978-1-0716-1464-8_10.

DOI:10.1007/978-1-0716-1464-8_10

PMID:34845689

Abstract

Northern blot analysis reveals information about RNA identity, size, and abundance. This technique has become an essential tool for the knowledge developed about viroids and also an excellent method for viroid detection. Here we describe the methodology of a Northern blot based in polyacrylamide gel electrophoresis under denaturing conditions, hybridized with a viroid full-length riboprobe labeled with chemiluminescence. Viroid detection with this approach entails positive signals, specific migration, and the differentiation of their circular and linear forms.

摘要

Northern blot 分析揭示了 RNA 的身份、大小和丰度信息。这项技术已成为了解类病毒的重要工具，也是检测类病毒的绝佳方法。在这里，我们描述了一种基于聚丙烯酰胺凝胶电泳在变性条件下进行的 Northern blot 方法，该方法与用化学发光标记的类病毒全长核糖探针杂交。使用这种方法进行类病毒检测需要出现阳性信号、特定的迁移以及区分其环状和线性形式。