The study of regulatory small RNAs, such as siRNAs and microRNAs in plants and other organisms, has necessitated techniques tailored to their unique features. Their analysis requires the use of sensitive and quantitative methods for their detection. The use of northern blot hybridization offers an attractive approach to address qualitative as well as quantitative features, and we describe here the use of radioactive and nonradioactive methods for signal detection. We highlight the advantages and shortcomings of these approaches and offer a detailed description of the techniques that best work for us, considering their use for the study of various small RNAs in multiple samples. We list relevant details and provide cautionary comments in those steps where we have detected potential difficulties.

错误:搜索内容不能为空,请输入英文关键词
错误:关键词超出字数限制,请精简
高级检索

Northern Blot Analysis of Plant MicroRNAs and Other Small RNAs

  • Yuniet Hernández,
  • Carlos De la Rosa,
  • José Luis Reyes

摘要

The study of regulatory small RNAs, such as siRNAs and microRNAs in plants and other organisms, has necessitated techniques tailored to their unique features. Their analysis requires the use of sensitive and quantitative methods for their detection. The use of northern blot hybridization offers an attractive approach to address qualitative as well as quantitative features, and we describe here the use of radioactive and nonradioactive methods for signal detection. We highlight the advantages and shortcomings of these approaches and offer a detailed description of the techniques that best work for us, considering their use for the study of various small RNAs in multiple samples. We list relevant details and provide cautionary comments in those steps where we have detected potential difficulties.