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The concentration of gel affects the resolution of DNA separation. The agarose gel is composed of microscopic pores through which the molecules travel, and there is an inverse relationship between the pore size of the agarose gel and the concentration – pore size decreases as the density of agarose fibers increases.
Agarose gel can have high gel strength at low concentration, making it suitable as an anti-convection medium for gel electrophoresis. Agarose gels as dilute as 0.15% can form slabs for gel electrophoresis. [12] The agarose polymer contains charged groups, in particular pyruvate and sulfate. [9]
For a standard agarose gel electrophoresis, 0.7% gel concentration gives good separation or resolution of large 5–10kb DNA fragments, while 2% gel concentration gives good resolution for small 0.2–1kb fragments.
Agarose gels do not have a uniform pore size, but are optimal for electrophoresis of proteins that are larger than 200 kDa. [10] Agarose gel electrophoresis can also be used for the separation of DNA fragments ranging from 50 base pair to several megabases (millions of bases), [11] the largest of which require specialized apparatus. The ...
Concentration Agarose concentration must be taken into account when selecting a marker. The gel percentage effects the migration of the DNA. [3] [6] Generally, the higher the gel concentration, the slower the rate at which the DNA will move through the gel. This is in addition to the role molecular weight plays in the migration of a DNA marker ...
The fluorescent dye is either present in the agarose gel, or is added in appropriate concentrations to the samples on the plastic film. A set of samples with known concentrations are spotted alongside the sample. The concentration of the unknown sample is then estimated by comparison with the fluorescence of these known concentrations.
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