Difference between revisions of "Loading dye"

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==Purpose==
 
==Purpose==
 
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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).
Loading dye is mix 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).
 
  
 
==Procurement==
 
==Procurement==

Revision as of 11:23, 25 August 2006

Purpose

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).

Procurement

Here's a sample recipe for loading dye.

  • Ficoll 400 (in 68-558F)
  • 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