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0.5M EDTA, pH 8.0
EDTA, 0.5 M, pH 8.0, is a sterile-filtered solution of 0.5 M EDTA in H2O treated with diethyl pyrocarbonate (DEPC).
Description
The loading dye increases the density of the sample and adds colours to the sample, thereby simplifying the loading process. The solution contains dyes that, in a electric field, move toward the anode at predictable rates.
In 1% agarose gels, bromophenol blue migrates with a 300 bp linear double-stranded DNA fragment, whereas xylene cyanol FF migrates at approximately the same rate as linear double-stranded DNA 4 kb length. These relationships are not significantly affected by the concentration (0.5 to 1.4%) of agarose in the gel.
Loading: The loading dye suits well for DNA samples dissolved either in water or in EDTA-containing buffer (as TE buffer). If DNA markers are not prediluted with the Loading dye solution, then mix. The loading buffer is 6x concentrated, that means you have to use it 5:1.
For DNA markers, apply 0.1 µg per 1 mm of agarose gel lane width. Often 1µg of marker is used in one electrophoresis run but it depends on the size of your gel and the comb.
Applications
Preparation of DNA ladders, markers, and samples for loading on agarose or polyacrylamide gels.
Recommended Storage Conditions: Store at room temperature or at 4 °C up to 12 months. For longer periods, store at -20 °C.
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The loading dye increases the density of the sample and adds colours to the sample, thereby simplifying the loading process. The solution contains dyes that, in a electric field, move toward the anode at predictable rates.