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Megaprime DNA Labeling System

Megaprime DNA标记系统

Company: Cytiva
Catalog#: RPN1604
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A High Resolution Short Interfering RNA (siRNA) Detection Method from Virus-infected Plants
[Abstract]  Plant viruses are strong inducers as well as targets of RNA silencing. In plants RNA silencing acts as a natural defense mechanism against viral infection and is associated with accumulation of virus-specific small interfering RNAs (siRNAs). The continuing discoveries, increasing awareness and interest in the regulatory roles of non-coding small RNAs have raised the need for methods that can reliably detect and quantitate the expression levels of small RNAs. Northern blot analysis of small RNAs involving the separation of RNA molecules using polyacrylamide gel electrophoresis (PAGE) has remained a popular and valuable analytical method to validate small RNAs. Northern blot analysis consist of resolving RNAs by gel electrophoresis, followed by transferring and fixing to nylon membranes as ... [摘要]  植物病毒是强诱导物以及RNA沉默的靶标。在植物中RNA沉默作为针对病毒感染的天然防御机制,并且与病毒特异性小干扰RNA(siRNA)的积累相关。持续的发现,增加对非编码小RNA的调节作用的意识和兴趣已经提出了对可以可靠地检测和定量小RNA的表达水平的方法的需要。涉及使用聚丙烯酰胺凝胶电泳(PAGE)分离RNA分子的小RNA的Northern印迹分析仍然是验证小RNA的普遍和有价值的分析方法。 Northern印迹分析由通过凝胶电泳分辨RNA,随后转移和固定到尼龙膜以及通过使用放射性探针的杂交检测组成。以下方案提供了用于从病毒感染的植物分离和检测小RNA的方法,并且成功地用于Panwar等人(2013a),Panwar等人( 2013b)。

32P Radioactive Probe Synthesis and Preparation
[Abstract]   To probe for a specific mRNA species by Northern blot, RNA from the agarose/formaldehyde gel needs to be transferred to a nylon membrane. RNA is detected by hybridization using a labeled probe. The probe is a DNA or RNA molecule that is chemically or radioactively labeled. In this protocol synthesis and preparation of a [32P]-dCTP-labeled probe is described. [摘要]  为了通过Northern印迹探测特定的mRNA种类,来自琼脂糖/甲醛凝胶的RNA需要转移到尼龙膜上。 使用标记的探针通过杂交来检测RNA。 探针是化学或放射性标记的DNA或RNA分子。 在该方案中,描述了[32 P] -dCTP标记的探针的合成和制备。