Alkaline Electrophoresis
Alkaline Electrophoresis - Related Videos
05:06
Alkaline Single Cell Gel Electrophoresis: A Sensitive Technique for Quantitative Estimation of DNA Damage in Individual Cancer Cells Following Chemotherapy
0 Views •
2025
In this video, we demonstrate an alkaline single-cell electrophoresis assay to quantify both single- and double-strand DNA breaks in doxorubicin-treated cells. Alkaline conditions disrupt the hydrogen bonds between DNA base pairs, resulting in the complete unwinding and separation of DNA strands. This allows differential migration of undamaged and damaged DNA fragments in agarose gel upon electrophoresis, revealing a comet-shaped structure of slowly migrating undamaged DNA forming the circular...
10:38
In Vivo Alkaline Comet Assay and Enzyme-modified Alkaline Comet Assay for Measuring DNA Strand Breaks and Oxidative DNA Damage in Rat Liver
0 Views •
Cited by 17 •
2016
The alkaline comet assay measures DNA strand breaks in eukaryotic cells. By adding an Endonuclease III or human 8-oxoguanine-DNA-N-glycosylase digestion step, the assay can efficiently detect oxidative DNA damage. We describe methods for using these assays to detect DNA damage in rat liver.
08:50
Capillary Electrophoresis (CE)
0 Views •
2023
Source: Laboratory of Dr. B. Jill Venton - University of Virginia Capillary electrophoresis (CE) is a separation technique that separates molecules in an electric field according to size and charge. CE is performed in a small glass tube called a capillary that is filled with an electrolyte solution. Analytes are separated due to differences in electrophoretic mobility, which varies with charge, solvent viscosity, and size. Traditional electrophoresis in gels is limited in the amount of voltage...
07:31
Two-Dimensional Gel Electrophoresis
0 Views •
2023
Two-dimensional gel electrophoresis (2DGE) is a technique that can resolve thousands of biomolecules from a mixture. This technique involves two distinct separation methods that have been coupled together: isoelectric focusing (IEF) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). This physically separates compounds across two axes of a gel by their isoelectric points (an electrochemical property) and their molecular weights. The procedure in this video covers the main...
09:22
DNA Gel Electrophoresis
0 Views •
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...
FAQs
The high-pH environment changes what the gel resolves by separating DNA strands before migration. This denaturation produces single-stranded molecules, so strand breaks and fragmentation can influence the pattern more directly than they would in an undisturbed double-stranded structure. That feature makes alkaline electrophoresis valuable when the research question concerns DNA integrity rather than only fragment separation.
Alkalinity is not merely a buffer condition; it is the mechanistic basis of the assay. By exposing DNA to high pH, the method converts the analyzed molecules into single strands, allowing structural damage to be examined in that state. Consequently, DNA damage that produces strand breaks or fragmentation can become evident in ways less apparent under neutral electrophoresis conditions.
Migration depends on both the porous gel and fragment length. The gel provides a medium through which charged nucleic-acid molecules move toward the oppositely charged electrode, while differently sized fragments can travel differently from one another. Interpreting the resulting separation therefore requires attention to fragment-size differences and the structural state created by alkaline treatment.
A supported workflow begins by placing the nucleic-acid sample in an alkaline buffer, allowing the high-pH condition to denature the DNA, and then examining migration through a porous gel under an electric field. The electrode arrangement drives movement of the charged molecules. This sequence links sample treatment with separation, so the observed pattern reflects both fragment length and DNA integrity.
Researchers apply alkaline electrophoresis to detect strand breaks and assess DNA fragmentation in nucleic-acid samples. These readouts are useful in molecular biology and genotoxicity studies because they provide evidence about whether DNA has retained its integrity. The method can therefore support investigations focused on structural changes in DNA and the extent of damage present in a sample.
In biological techniques, the method is especially relevant to studies of genome stability, cellular stress, and repair mechanisms. Its value lies in connecting a separation pattern with possible DNA damage: fragmentation or strand breaks become important features for interpreting sample condition. Because alkaline conditions can reveal damage less apparent at neutral pH, the approach adds information beyond neutral analysis.