Hybrid Purification
Hybrid Purification - Related Videos
08:16
Plasmid Purification
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2023
Plasmid purification is a technique used to isolate and purify plasmid DNA from genomic DNA, proteins, ribosomes, and the bacterial cell wall. A plasmid is a small, circular, double-stranded DNA that is used as a carrier of specific DNA molecules. When introduced into a host organism via transformation, a plasmid will be replicated, creating numerous copies of the DNA fragment under study. In this video, a step-by-step generalized procedure is described for how to perform plasmid purification.
06:30
Gel Purification
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2023
Gel purification is used to recover DNA fragments after electrophoretic separation. DNA recovery from an agarose gel includes three basic steps: binding, washing and eluting from a silica column. DNA is believed to bind to silica in the presence of high salt via a salt bridge. Following binding, DNA is washed of impurities and eluted under low salt conditions disrupting this interaction. This video goes through a step-by-step, generalized procedure for cutting out a band from the gel, gel...
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.
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 selectivity comes from complementary base pairing between the probe and the desired DNA or RNA sequence. Under controlled hybridization conditions, matching targets form stable nucleic acid–probe hybrids, whereas unrelated molecules bind less effectively or remain unbound. This molecular recognition allows the method to distinguish a selected genetic sequence within a complex biological mixture.
Affinity supports, including immobilized oligonucleotides and magnetic beads, provide a way to recover probe-bound targets from the sample. Washing removes nonspecific molecules and weakly associated material while retaining the desired hybrids. The balance between hybrid stability and washing stringency therefore affects both enrichment quality and the amount of background carried into subsequent genetic analysis.
Hybrid Purification enriches molecules that already contain a chosen sequence, while amplification is performed afterward when additional copies are needed for analysis. Purification reduces competing background before amplification, sequencing, or detection rather than replacing those processes. Combining selective enrichment with downstream analysis can focus measurements on rare transcripts, genomic regions, or pathogen sequences.
A typical workflow begins with a complex biological nucleic acid mixture and sequence-specific probes designed for the target. Hybridization is allowed under controlled conditions, after which probe-bound material is captured through an affinity support. Washing removes nonspecific molecules, and the recovered target-enriched fraction proceeds to amplification, sequencing, or another molecular analysis.
The approach is useful when a sequence of interest represents only a small fraction of the starting material or when unrelated nucleic acids create substantial background. It can enrich rare transcripts, selected genomic regions, and pathogen sequences before sensitive genetic detection. Focusing the sample in this way supports more targeted analysis than examining the original mixture alone.
The principal outcome is a recovered fraction with a higher concentration of the selected nucleic acid and less nonspecific material. This improved target-to-background relationship can support sensitive detection and more focused molecular analysis. In genetics, the enriched material may be directed toward amplification, sequencing, or other assays that require clearer access to a particular sequence.