Northern Hybridization
Northern Hybridization - Related Videos
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.
03:35
Hybridization of Atomic Orbitals II
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2020
sp3d and sp3d 2 Hybridization To describe the five bonding orbitals in a trigonal bipyramidal arrangement, we must use five of the valence shell atomic orbitals (the s orbital, the three p orbitals, and one of the d orbitals), which gives five sp3d hybrid orbitals. With an octahedral arrangement of six hybrid orbitals, we must use six valence shell atomic orbitals (the s orbital, the three p orbitals, and two of the d orbitals in its valence shell), which gives six sp3d 2 hybrid orbitals.
03:24
Hybridization of Atomic Orbitals I
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2020
The mathematical expression known as the wave function, ψ, contains information about each orbital and the wavelike properties of electrons in an isolated atom. When atoms are bound together in a molecule, the wave functions combine to produce new mathematical descriptions that have different shapes. This process of combining the wave functions for atomic orbitals is called hybridization and is mathematically accomplished by the linear combination of atomic orbitals. The new orbitals that...
08:00
Whole-Mount In Situ Hybridization
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2023
Whole-mount in situ hybridization (WMISH) is a common technique used for visualizing the location of expressed RNAs in embryos. In this process, synthetically produced RNA probes are first complementarily bound, or "hybridized," to the transcripts of target genes. Immunohistochemistry or fluorescence is then used to detect these RNA hybrids, revealing spatial and temporal patterns of gene expression. Unlike traditional in situ hybridization techniques, which require thin tissue sections whose...
FAQs
Probe complementarity determines detection specificity: a labeled nucleic acid probe binds more effectively when its sequence matches the target RNA. The hybridization conditions are important because they promote specific base pairing rather than indiscriminate association. This sequence-based recognition lets investigators distinguish a chosen transcript from other RNA molecules in the sample.
Gel electrophoresis separates RNA molecules according to size before detection. After the RNA reaches the membrane, the position of a probe-generated signal identifies the approximate size of the recognized transcript. Comparing signal positions can therefore reveal transcript-size differences and contribute to analysis of transcript structure, not merely whether RNA is present.
The labeled probe converts sequence recognition into a detectable result. When complementary base pairing occurs under conditions that favor specificity, the probe marks the target RNA on the membrane. The resulting signal supports two related judgments: whether the transcript is detectable and its approximate abundance, while the signal location supplies size information.
A typical setup requires an RNA sample, a gel for size separation, a membrane for transfer, and a labeled nucleic acid probe. The hybridization stage must provide conditions that promote specific base pairing, because probe recognition depends on sequence complementarity and affects the interpretability of the resulting signal.
It can compare RNA detection patterns across developmental stages, tissues, or experimental and environmental conditions. Such comparisons connect a transcript with regulated gene expression, including tissue-specific or developmentally controlled changes. Because the method also reports transcript size, the same experiment can relate expression differences to transcript structure.
Results are interpreted by considering both signal detection and signal position. Detection indicates transcript presence, approximate signal abundance supports comparisons, and position indicates transcript size. Together, these observations help examine RNA processing and transcript structure alongside gene expression, allowing investigators to assess whether conditions affect transcript levels, transcript size, or both.