Gel Electrophoresis Simulator: DNA Fragment Separation & Band Migration

simulator beginner ~8 min
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Migration = 42 mm — 1 kb fragment, 1% gel, 100V, 45 min

A 1000 bp DNA fragment migrates approximately 42 mm in a 1% agarose gel at 100V over 45 minutes, well-separated from common ladder markers.

Formula

μ = v/E = d/(t·E) (electrophoretic mobility)
d ∝ -log₁₀(bp) (Ferguson plot relationship)
E = V/L (electric field strength, V/cm)

Separation by Size

Gel electrophoresis is the workhorse technique of molecular biology — virtually every genetics lab runs gels daily to verify PCR products, check restriction digests, and analyze DNA quality. The principle is elegant: DNA's uniform negative charge drives it through a porous agarose matrix under an electric field, with smaller fragments migrating faster than larger ones. The result is a pattern of discrete bands, each representing fragments of a specific size, that can be visualized with fluorescent dyes.

The Agarose Matrix

Agarose — a polysaccharide extracted from seaweed — forms a gel with pore sizes determined by concentration. At 0.5%, pores are large enough for fragments up to 30 kb to migrate. At 2%, the tight network resolves fragments as small as 100 bp but traps anything larger than 2 kb. Choosing the right concentration is the first decision in any electrophoresis experiment. This simulation models the pore network and shows how fragment size interacts with gel concentration to determine migration distance.

Voltage, Time & Resolution

Migration distance increases with both voltage and time, but these parameters also affect band sharpness. High voltage causes heating (Joule effect) that can melt the gel and cause band smearing. The optimal field strength is 5-8 V/cm for standard applications. Longer run times improve separation between similarly-sized fragments but risk running small fragments off the gel. The simulation lets you balance these tradeoffs and see how different conditions affect both migration distance and band resolution.

Reading the Gel

Interpreting a gel image is a fundamental skill in molecular biology. Sharp, bright bands indicate clean, concentrated DNA of uniform size. Smearing suggests degradation. Multiple unexpected bands may indicate non-specific amplification or star activity in restriction digests. The DNA ladder provides the size reference — plotting log(size) versus migration distance yields a straight line (Ferguson plot) from which unknown fragment sizes can be interpolated. This simulation includes a standard ladder lane for comparison.

FAQ

How does gel electrophoresis separate DNA?

DNA carries a uniform negative charge from its phosphate backbone, so it migrates toward the positive electrode in an electric field. Smaller fragments snake through the agarose pore network faster than larger ones, creating size-based separation. Migration distance is inversely proportional to the log of fragment size — a linear relationship on semi-log plots.

How does agarose concentration affect separation?

Higher agarose concentration creates smaller pores, improving resolution of small fragments but slowing or preventing migration of large ones. 0.5% agarose resolves 1-30 kb fragments, 1% resolves 0.5-10 kb, and 2% resolves 0.1-2 kb. Polyacrylamide gels are used for even finer resolution below 500 bp.

What is a DNA ladder and why is it used?

A DNA ladder is a mixture of DNA fragments of known sizes, loaded alongside samples as a reference standard. By comparing sample band positions to the ladder, you can estimate fragment sizes. Common ladders include the 1 kb ladder (0.5-10 kb range) and 100 bp ladder (100-1500 bp range).

How do you visualize DNA bands?

DNA bands are typically visualized using fluorescent intercalating dyes. Ethidium bromide (UV-activated, mutagenic) was the traditional choice. Safer alternatives include SYBR Safe, GelRed, and GelGreen that fluoresce under blue light. The fluorescence intensity is proportional to DNA mass, enabling semi-quantitative analysis.

Sources

Embed

<iframe src="https://homo-deus.com/lab/biotechnology/gel-electrophoresis/embed" width="100%" height="400" frameborder="0"></iframe>
View source on GitHub