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FUNDAMENTAL PRINCIPLES OF ELECTROPHORESIS Articles

Faint Bands, High Background

By National Diagnostics | August 23, 2013 | Comments Off on Faint Bands, High Background

A high background which obscures the bands, or in combination with fainter than usual bands, indicates dye binding to the gel matrix, or contamination of the matrix with a dye-binding material (most often a protein).…

Faint bands, low background

By National Diagnostics | August 16, 2013 | Comments Off on Faint bands, low background

SDS in the staining solution:  Most users save and re-use their staining solutions.  This is fine for a couple of cycles, but over time enough SDS elutes from the gels to accumulate to significant levels…

Coomassie Blue Stain- Troubleshooting

By National Diagnostics | August 12, 2013 | Comments Off on Coomassie Blue Stain- Troubleshooting

If your gel doesn’t look like this one, click on the problem below to find the solution: Faint bands on a low background Learn More Faint bands on a high background Learn More Uneven Staining…

Radioactive Emissions and the Use of Isotopes in Research

By National Diagnostics | October 10, 2011 | Comments Off on Radioactive Emissions and the Use of Isotopes in Research

Radioactive decay occurs with the emission of particles or electromagnetic radiation from an atom due to a change within its nucleus. Forms of radioactive emission include alpha particles (α), beta particles (β), and gamma rays…

Ethidium Bromide Staining

By National Diagnostics | September 12, 2011 | Comments Off on Ethidium Bromide Staining

Bands in gels stained with ethidium bromide fluoresce under ultraviolet light. The most commonly used stain for detecting DNA/RNA is ethidium bromide. Ethidium bromide is a DNA interchelator, inserting itself into the spaces between the…

Horizontal and Vertical Gel Systems – Vertical Tube Gels

By National Diagnostics | August 15, 2011 | Comments Off on Horizontal and Vertical Gel Systems – Vertical Tube Gels

Tube gels were used frequently in the development of gel electrophoresis. Although they are still used for some applications (most notably for isoelectric focusing as part of 2D electrophoresis), tube gels have been superseded by…

Horizontal and Vertical Gel Systems – The Vertical Slab Gel System

By National Diagnostics | August 15, 2011 | Comments Off on Horizontal and Vertical Gel Systems – The Vertical Slab Gel System

A typical vertical apparatus used for sequencing is shown in the figure below. This system shows the components common to all vertical slab systems. The gel is cast between two glass plates, separated by spacers,…

Horizontal and Vertical Gel Systems – The Horizontal Gel System

By National Diagnostics | August 15, 2011 | Comments Off on Horizontal and Vertical Gel Systems – The Horizontal Gel System

A gel electrophoresis apparatus must allow the researcher to maintain a uniform electric field across the gel, provide cooling to prevent thermal artifacts, and allow access to the gel for sample loading and monitoring the…

Buffer Additives-Reducing Agents

By National Diagnostics | August 12, 2011 | Comments Off on Buffer Additives-Reducing Agents

Disulfide bonds between or within sample protein molecules can lead to the formation of aggregates as well as play a role in the binding of the subunits of many proteins. It is usually desirable to…

Buffer Additives-Surfactants

By National Diagnostics | August 12, 2011 | Comments Off on Buffer Additives-Surfactants

A crucial initial step in the electrophoretic separation of proteins is the solubilization of the sample molecules. This is especially true if there are extensive nonpolar interactions. Although urea in high concentration was often employed…