Agarose gel loading buffer: Difference between revisions
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===Ficoll & Orange G === | ===Ficoll & Orange G === | ||
*Ficoll 400 | *Ficoll 400 | ||
*Deionized water | *Deionized water | ||
*Orange G dye | *Orange G dye | ||
Revision as of 05:11, 20 April 2007
Loading dye is mixed with DNA samples for use in agarose gel electrophoresis. It generally contains a dye to assess how "fast" your gel is running and a reagent to render your samples denser than the running buffer (so that the samples sink in the well).
Material
Density
- Ficoll
- sucrose
Colour
- xylene cyanol (Sigma X4126)
- bromophenol blue (Sigma B8026)
- Orange G
Recipes
Ficoll & Orange G
- Ficoll 400
- Deionized water
- Orange G dye
Dissolve 1.5 g of Ficoll in 10 mL of deionized water. Add very small amounts of Orange G dye such that the loading dye is dark orange. Store in small aliquots at 4°C (room temperature is okay too). To use, add and mix 1/5th volume of loading dye to DNA solutions prior to loading into the wells of gels.
Specific recipes
Use
- Orange G: generally runs very fast (<100 bp)
- Bromophenol blue: purple, generally runs at ~500bp (depending on percentage agarose)
- Xylene cyanol: blue, runs at ~4kb
