Related Experiment Video
Updated: Aug 13, 2025
14:41
RNA Blot Analysis for the Detection and Quantification of Plant MicroRNAs
Published on: July 11, 2020
10.7K
Detection of MicroRNAs by Northern Blot
Claudia Martinho1,2, Sara Lopez-Gomollon3
1Department of Plant Sciences, University of Cambridge, Cambridge, UK.
Methods in Molecular Biology (Clifton, N.J.)
|January 23, 2023
Summary
Northern blotting is a quantitative method to analyze small RNAs (sRNAs), including microRNAs (miRNAs), small interfering RNAs (siRNAs), and PIWI-interacting RNAs (piRNAs). This guide details protocols for radioactive and non-radioactive sRNA Northern blot analysis.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Small RNAs (sRNAs) regulate gene expression by interacting with complementary RNA targets.
- Key sRNA classes include microRNAs (miRNAs), small interfering RNAs (siRNAs), and PIWI-interacting RNAs (piRNAs).
- The expression patterns of sRNAs are crucial for their biological functions.
Purpose of the Study:
- To provide detailed protocols for Northern blot analysis of small RNAs (sRNAs).
- To describe both radioactive and non-radioactive methods for sRNA detection.
- To offer practical guidance, including reagent preparation and commercial alternatives.
Main Methods:
- RNA extraction from biological samples.
- Polyacrylamide gel electrophoresis (PAGE) for sRNA separation.
- Membrane transfer, hybridization, and detection using oligonucleotide probes.
Main Results:
- Established protocols for radioactive and non-radioactive sRNA Northern blotting.
- Included methods for reagent preparation and highlighted commercial alternatives.
- Provided practical tips and experienced-based suggestions for successful analysis.
Conclusions:
- Northern blotting is a reliable and accessible technique for quantitative sRNA analysis.
- The provided protocols facilitate the study of sRNA abundance and expression.
- This guide supports researchers in investigating the roles of various sRNA classes.
