Hybridization Ligation
Hybridization Ligation - Related Videos
07:34
DNA Ligation Reactions
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2023
In molecular biology, ligation refers to the joining of two DNA fragments through the formation of a phosphodiester bond. An enzyme known as a ligase catalyzes the ligation reaction. In the cell, ligases repair single and double strand breaks that occur during DNA replication. In the laboratory, DNA ligase is used during molecular cloning to join DNA fragments of inserts with vectors – carrier DNA molecules that will replicate target fragments in host organisms. This video provides an...
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Cecal Ligation Puncture Procedure
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The mouse model of cecal ligation and puncture as a valuable tool for the study of human sepsis.
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Proximity Ligation Assay to Study In Situ Protein-Protein Interactions
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2025
This video describes the proximity ligation assay to study in situ protein-protein interactions in nematode cells. The close proximity of the proteins of interest generates the circular DNA template with the help of a PLA probe attached to a secondary antibody. The probe is further amplified using the amplification solution, which produces a long single-stranded DNA followed by the hybridization of the detector probe, enabling the localization of protein-protein interactions in cells.
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Modified Technique for Coronary Artery Ligation in Mice
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The surgical procedure used to induce experimental myocardial infarction in mice begins with left thoracotomy between the third and the fourth ribs in order to visualize the anterior surface of the heart and left lung. The left coronary artery is ligated, the chest is closed and the mouse is allowed to recover spontaneously.
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
Adjacency places the two probe ends directly beside one another on the same DNA or RNA strand. This alignment creates the substrate that DNA ligase can seal. If the probes bind at separated positions, their ends are not properly positioned for joining, so the assay does not efficiently generate the ligated product needed for detection or later analysis.
Specificity arises from two linked requirements: complementary nucleic acid pairing and successful enzymatic joining. Correctly matched probes hybridize beside each other and present aligned ends for ligase. A sequence difference can interfere with this recognition or the joining step, allowing the method to distinguish particular DNA or RNA sequences, including variants used in mutation or genotype analysis.
DNA ligase converts probe alignment into a covalently joined product by sealing the break between adjacent probe ends. Hybridization alone indicates that probes can pair with a target, whereas ligation records that the probes were positioned appropriately for enzymatic joining. The resulting product can then support downstream detection, amplification, or sequence analysis.
Hybridization alone depends on complementary pairing between a probe and its target. Hybridization ligation adds an enzymatic joining requirement between probes bound next to each other, creating an additional sequence-dependent checkpoint. This extra step can improve discrimination among closely related sequences and produces a ligated product that is suitable for subsequent detection or amplification.
The workflow begins by bringing oligonucleotide probes and the DNA or RNA target together so the probes can hybridize to adjacent complementary regions. DNA ligase is then used to seal the aligned probe ends when the sequence arrangement is correct. The ligation product is subsequently detected, amplified, or analyzed according to the experimental objective.
The method is useful when an experiment must distinguish specific sequence alternatives rather than merely detect related nucleic acids. Probes are designed to recognize the relevant target regions, and sequence-dependent ligation indicates whether the expected arrangement is present. This supports mutation analysis and genotype discrimination by converting precise recognition into a product that can be examined.
For pathogen identification, probes can target characteristic DNA or RNA sequences, allowing ligation to signal recognition of the selected molecular signature. In gene analysis, the same principle supports precise examination of chosen sequences. Because the joined products can undergo downstream detection, amplification, or analysis, the approach is adaptable to studies requiring sequence-specific molecular identification.