Northern Analysis
Northern Analysis - Related Videos
09:39
Enhanced Northern Blot Detection of Small RNA Species in Drosophila Melanogaster
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Cited by 5 •
2014
The aim of this publication is to visualize and discuss the operative steps of an Enhanced Northern Blot protocol on RNA extracted from Drosophila melanogaster embryos, cells, and tissues. This protocol is particularly useful for the efficient detection of small RNA species.
04:33
Gel Electrophoresis and Northern Blotting to Detect tRNA Charging Levels in E. coli
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2025
Source: Stenum, T. S., et al. Quantification of the Abundance and Charging Levels of Transfer RNAs in Escherichia coli. J. Vis. Exp. (2017)This video demonstrates the use of gel electrophoresis and northern blotting to detect and compare the charging levels of transfer RNAs in E. coli by separating aminoacylated and deacylated forms.
08:51
Northern Blot to Detect and Quantify Specific MicroRNA in Total RNA Extract of Plant Tissue
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2025
In this video, we demonstrate the northern blot, a hybridization technique to detect and quantify microRNAs (miRNAs) from total RNA extracted from plant tissue.
15:04
A Neuronal and Astrocyte Co-Culture Assay for High Content Analysis of Neurotoxicity
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Cited by 46 •
2009
This article describes a novel protocol and reagent set designed for sensitive measurement of neurotoxic effects of compounds and treatments on co-cultures of neurons and astrocytes using high content analysis. Results demonstrate that high content analysis represents an exciting novel technology for neurotoxicity assessment.
10:39
Automated Analysis of Intracellular Phenotypes of Salmonella Using ImageJ
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Cited by 3 •
2022
Salmonella invades and replicates inside intestinal epithelial cells both in Salmonella-specific vacuoles and free in the cytosol (hyper-replication). A high-throughput fluorescence microscopy-based protocol is described here to quantify the intracellular phenotypes of Salmonella by two complementary image analyses through ImageJ, reaching single-cell resolution and scoring.
FAQs
Specificity comes from base pairing between the labeled nucleic acid probe and a complementary RNA sequence on the membrane. Only transcripts carrying the matching sequence are visualized as bands, so the probe determines which RNA species the assay can recognize. This makes Northern analysis useful when the question concerns one transcript or a defined related set rather than total RNA.
Gel electrophoresis contributes size information before detection. RNA molecules migrate through the gel according to their size, and their positions are preserved after transfer to the membrane. A detected band therefore supports two observations at once: whether the target transcript is present and its approximate length. Band intensity can also be compared across samples as an indicator of relative abundance.
Transcript-size differences can be biologically informative rather than merely technical variation. When samples show bands at different approximate lengths, the pattern may be associated with alternative RNA processing or changes in gene regulation. Comparing these patterns across experimental groups can therefore distinguish a simple change in transcript amount from a change in the RNA species being produced.
An analysis proceeds through a linked sequence of steps: RNA is first separated by size in a gel, then transferred to a membrane, and finally exposed to a labeled nucleic acid probe. Hybridization identifies the complementary sequence, while the resulting band pattern is interpreted for transcript presence, relative abundance, and approximate length. Each stage contributes a different kind of information.
Comparative studies are a central application. Researchers can examine the same RNA target across tissues, developmental stages, or experimental conditions, then compare band presence, approximate size, and relative abundance. This design links molecular measurements to biological context: a tissue-restricted band suggests differing expression, whereas a changed band size points toward transcript-size variation that may accompany regulation or processing.
Northern analysis is especially useful as a validation tool when an experiment has identified an RNA expression pattern that needs direct transcript-level confirmation. Its bands can show whether the expected RNA is present, how its approximate length compares among samples, and whether abundance changes track the proposed pattern. Newer methods may provide greater throughput, but this approach retains interpretive value for focused comparisons.