Wide Combed Gel
Wide Combed Gel - Related Videos
09:22
DNA Gel Electrophoresis
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2023
DNA gel electrophoresis is a technique used for the detection and separation of DNA molecules. An electric field is applied to a gel matrix comprised of agarose, and within the gel, charge particles will migrate and separate based on size. The negatively charged phosphates of the DNA backbone cause DNA fragments to move toward the anode - a positively charged electrode. The video explains the mechanism by which DNA fragments are resolved on an agarose gel, and it provides a step-by-step...
11:03
Pouring and Running a Protein Gel by reusing Commercial Cassettes
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Cited by 2 •
2012
Our protocol demonstrates how to pour multiple protein gels at a time by recycling Invitrogen Nupage Novex minigel cassettes, and inexpensive materials purchased at a home improvement store. This economical and streamlined method includes a way to store the gels at 4°C for a few weeks. By re-using the plastic gel cassettes from commercially available gels, labs that run frequent protein gels can save significant costs and help the environment.
07:10
Agarose Gel Electrophoresis for the Separation of DNA Fragments
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Cited by 599 •
2012
A basic protocol for the separation of DNA fragments using agarose gel electrophoresis is described.
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...
09:24
Mucin Agarose Gel Electrophoresis: Western Blotting for High-molecular-weight Glycoproteins
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Cited by 32 •
2016
Mucins are high-molecular-weight glycoconjugates, with size ranging from 0.2 to 200 megadalton (MDa). As a result of their size, mucins do not penetrate conventional polyacrylamide gels and require larger pores for separation. We provide a detailed protocol for mucin agarose gel electrophoresis to assess relative quantification and study polymer assembly.
FAQs
Broad wells allow researchers to load larger sample volumes than standard wells, which is important when a preparation contains limited concentration but a substantial total amount of DNA. This capacity supports side-by-side comparison of samples and provides enough separated material for later recovery. The format therefore connects analytical separation with preparative work.
An applied electric field drives nucleic acids through the agarose matrix toward the oppositely charged electrode. Molecules separate primarily according to size as they migrate, so the wide comb changes the loading capacity rather than replacing the underlying electrophoretic mechanism. Researchers can therefore use the same size-based separation principle with larger loaded quantities.
The format is preferable when researchers need to accommodate more sample per well or recover relatively large quantities of nucleic acid after separation. Standard systems remain suitable when ordinary loading capacity is sufficient, whereas broad wells are particularly useful for workflows that combine band resolution with purification, downstream analysis, or molecular cloning.
A typical workflow uses an agarose matrix cast with a wide comb, followed by loading the nucleic acid samples into its broad wells. Researchers then apply an electric field so the molecules migrate and separate by size. After resolving the bands, selected material can be recovered for purification, downstream analysis, or cloning.
Separated bands allow researchers to resolve and compare nucleic acid samples according to their migration through the agarose matrix. The resulting pattern can support assessment of relative fragment sizes and selection of material for recovery. Because the wells accept larger volumes, the same run can also generate more nucleic acid for subsequent work.
In preparative workflows, researchers load relatively large quantities of nucleic acid, separate the material in agarose, and recover a selected product or band. The recovered nucleic acid can then undergo purification, downstream analysis, or molecular cloning. This makes the format useful when electrophoresis must provide material, not merely a visual comparison.