Southern Hybridization
Southern Hybridization - Related Videos
05:42
Southern Blotting to Detect Homologous Recombination in the Mouse Embryonic Stem Cell Genome
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2025
In this video, we describe the Southern blotting technique to identify homologous recombination events in the genome of mouse embryonic stem cells using radiolabeled probes. DNA regions containing homologous recombination events appear as distinct bands when the membrane containing the probe-bound DNA is exposed to X-rays.
02:57
Southern Blot
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2021
Agarose gel electrophoresis is very useful in separating DNA fragments by size. Running a DNA ladder containing fragments of the known length alongside the sample helps determine the approximate length of the sample DNA fragments. However, additional steps are needed to verify the sequence identity of the sample DNA fragments. Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...
06:34
A Component-resolved Diagnostic Approach for a Study on Grass Pollen Allergens in Chinese Southerners with Allergic Rhinitis and/or Asthma
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Cited by 4 •
2017
This work describes a protocol that uses a component-resolved approach to study sensitization to grass pollen allergens in a cohort of patients from southern China with allergic rhinitis and/or asthma.
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...
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.
FAQs
Restriction-enzyme digestion creates DNA fragments that can be resolved by size during gel electrophoresis, giving the probe a set of separated targets to interrogate. After transfer to a membrane, the labeled complementary probe binds the matching sequence under controlled conditions. The resulting signal therefore reflects both sequence complementarity and the fragment pattern produced by digestion.
The probe provides sequence-level specificity in a complex genome. Because it is complementary to the DNA region of interest, it can distinguish the target from unrelated fragments after transfer to the membrane. Labeling makes the bound probe detectable, allowing investigators to locate the target and examine how its position among DNA fragments may reflect genome organization.
Compared with newer nucleic acid amplification and sequencing methods, Southern hybridization is generally slower, but it answers a different structural question. Its value lies in confirming genomic structure and examining stable genetic changes. This distinction supports its continued use when researchers need information about sequence organization or integration rather than only a faster molecular analysis.
Probe binding depends on controlled hybridization conditions, so this stage is central to obtaining an interpretable target signal. The workflow also relies on maintaining the relationship between restriction-digested fragments, their separation by gel electrophoresis, and their transfer to a membrane. Together, these conditions allow the labeled probe to be evaluated against a defined fragment pattern.
In infection studies, the method can assess whether pathogen DNA is integrated into genomic material and examine the organization of that DNA. Because the analysis considers target-containing fragments within a larger genome, it can provide structural evidence that complements simple detection of pathogen sequences. This makes it relevant to questions about stable genetic changes.
Applied to immunology, Southern hybridization can analyze rearrangements in immune receptor loci. The relevant question is not merely whether DNA is present, but how target sequences are organized within restriction-generated fragments. This use connects a DNA-level pattern to changes in immune receptor gene organization, providing a molecular approach for examining rearranged loci.