Hybridization Assay
Hybridization Assay - Related Videos
05:20
Yeast Two-Hybrid Assay to Determine Protein Self-Association in Yeast Cells
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2025
This video demonstrates the yeast two-hybrid assay to detect specific protein self-interactions. In this assay, the protein of interest is tagged to two domains of a transcription factor that encodes the β-galactosidase enzyme. When the two proteins interact, this causes transcription and translation to form the enzyme, which is then detected using a biochemical assay.
10:15
Multiplex PCR and Reverse Line Blot Hybridization Assay (mPCR/RLB)
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Cited by 21 •
2011
An inexpensive, high throughput method for simultaneous detection of up to 43 molecular targets is described. Applications of mPCR/RLB include microbial typing and detection of multiple pathogens from clinical samples.
04:12
Modified Yeast-One Hybrid Assay to Detect Heteromeric Protein Complex-DNA Interactions
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2025
In this video, we demonstrate the modified yeast one-hybrid assay, which detects multiple proteins interacting with target DNA. The transformed yeast cells contain the target DNA sequence upstream of the reporter gene. Additionally, they express transcription factor proteins. When the proteins form complexes and bind to the target DNA, the reporter gene is expressed and can be detected enzymatically.
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
These variables set the assay’s stringency, meaning how strictly probe and target strands must match to remain associated. Temperature, salt concentration, and washing conditions work together: appropriate settings preserve binding from complementary sequences while removing nonmatching strands. Changing stringency therefore affects specificity and helps determine whether a detected signal reflects the intended nucleic acid target.
Probe complementarity determines which sequence can be recognized. A DNA or RNA probe is selected for correspondence with the target, so base pairing favors the intended strand over unrelated nucleic acids. This sequence-level selectivity is especially useful when infection studies must identify a pathogen genome or discriminate between closely related strains.
Closely related strains may share much sequence but differ at particular target sequences. Probes directed at those distinguishing regions can produce different binding outcomes under controlled assay conditions. Comparing the resulting signals allows investigators to identify the relevant genome or separate strains, provided the probe-target match and washing conditions preserve the intended specificity.
A basic workflow brings the nucleic-acid sample into contact with a complementary DNA or RNA probe, allows annealing under controlled temperature and salt conditions, and washes away strands that do not remain associated. The assay then uses the probe’s detectable label to reveal the target-associated signal. These steps connect molecular recognition to an interpretable result.
Signal strength provides an indication of how much target is present because the assay’s principle links detectable output to target abundance. In infection research, this supports measurements of pathogen nucleic acids as well as host or microbial gene expression. Interpretation is strongest when samples are examined under comparable assay conditions, since temperature, salt, and washing affect retained binding.
Hybridization Assay supports diagnostic testing by revealing whether a characteristic pathogen genome is present. The same sequence-specific readout can be applied across samples for surveillance, helping investigators track the occurrence of pathogen nucleic acids and distinguish related strains. Its value comes from combining a recognizable molecular target with controlled removal of nonspecific binding.
In immunology and infection studies, the method can examine both sides of the host-pathogen interaction. Probes may target pathogen genomes to identify the infectious agent, or host and microbial sequences to measure gene expression. Those measurements help relate infection to altered cellular responses, linking sequence-level detection with broader immune and infection biology.