Gel Plug Isolation
Gel Plug Isolation - Related Videos
08:16
In Vivo Study of Human Endothelial-Pericyte Interaction Using the Matrix Gel Plug Assay in Mouse
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Cited by 11 •
2016
We present a protocol to study human endothelial-pericyte interactions in mouse using a variation of the matrix gel plug angiogenesis assay.
09:03
Modified In Vivo Matrix Gel Plug Assay for Angiogenesis Studies
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2023
The method presented here can evaluate the effect of reagents on angiogenesis or vascular permeability in vivo without staining. The method uses dextran-FITC injection via the tail vein to visualize neo-vessels or vascular leakage.
01:22
Formation of the Platelet Plug
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2024
The platelet phase, the second stage of hemostasis, commences around 15-20 seconds after an injury. It follows and overlaps with the vascular phase, during which blood vessels constrict to minimize blood loss. As the injured blood vessel contracts, endothelial cells undergo contraction, revealing collagen fibers in the basement membrane and underlying connective tissue. Furthermore, the plasma membrane of endothelial cells becomes adhesive, preparing the site for platelet adhesion. Platelets...
03:44
Isolation and Visualization of Rotavirus Genome via Gel Electrophoresis
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2026
Source: Philip, A. A., et al., Simplified Reverse Genetics Method to Recover Recombinant Rotaviruses Expressing Reporter Proteins. J. Vis. Exp. (2020)This video demonstrates the isolation of rotavirus double-stranded RNA from clarified cell lysates. It further illustrates the separation and visualization of genome segments through polyacrylamide gel electrophoresis.
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...
FAQs
Once the molten agarose cools, it forms a porous solid network that keeps the biological material in place. Reagents can move through this network for washing, lysis, and protein removal, while the embedded DNA remains physically supported. This arrangement reduces the need to transfer or manipulate long DNA molecules, helping preserve sample integrity during preparation.
Detergents disrupt cellular structures, allowing intracellular contents to become accessible, while enzymes help break down proteins that could remain associated with the sample. Because these treatments occur within the gel plug, the DNA stays embedded as cellular components are removed. The combined treatment prepares cleaner genomic material for later molecular analyses.
Very large DNA molecules are vulnerable to mechanical breakage during pipetting, transfers, and other handling steps. Immobilization within agarose limits those movements and therefore supports recovery of intact, high-molecular-weight DNA. Preserving long DNA molecules is particularly important when researchers need genomic preparations suitable for restriction profiling or pulsed-field gel electrophoresis.
After cellular lysis and protein removal, the retained genomic DNA can be used in analyses that distinguish biological samples by their DNA patterns. Restriction profiling examines fragments produced from the prepared DNA, whereas pulsed-field gel electrophoresis supports separation and comparison of large DNA fragments. These outputs can contribute to pathogen and strain characterization in infection research.
The sample is first suspended in molten agarose, and the mixture is cooled so the agarose solidifies around the cells or microorganisms. The resulting plug can then be exposed to detergents and enzymes for lysis and protein removal, followed by washing and further processing. This workflow keeps the sample contained while preparing its DNA for downstream analysis.
The method is useful when researchers need to isolate pathogens, prepare genomic DNA, or compare strains while protecting fragile or very large molecules. Its contained processing format supports sample integrity through lysis and washing, making it relevant to downstream restriction profiling and pulsed-field gel electrophoresis. These applications connect physical sample preservation with infection-focused genomic characterization.