Lithium electrophoresis buffer - Why does it not work? (Feb/03/2011 )
I have been reading the research by Brody and Kern (2004) about using a sodium boric acid buffer and a lithium borate buffer for gel electrophoresis. I firstly tried the sodium buffer and the results were reasonable, but not perfect. I then tried the lithium buffer expecting better results due to research suggesting that it is one of the best buffers and also the positive feedback that people have given on its performance.
However, when I tried 1mM lithium borate, the results were terrible. I was using a 1.7% gel and the bands were very smeared. I tried the buffer twice using different voltages (90V and 280V), and the results were pretty much the same.
Any insight as to why this is happening, or what I am doing wrong? Thanks.
Baileys on Thu Feb 3 17:59:19 2011 said:
I have been reading the research by Brody and Kern (2004) about using a sodium boric acid buffer and a lithium borate buffer for gel electrophoresis. I firstly tried the sodium buffer and the results were reasonable, but not perfect. I then tried the lithium buffer expecting better results due to research suggesting that it is one of the best buffers and also the positive feedback that people have given on its performance.
However, when I tried 1mM lithium borate, the results were terrible. I was using a 1.7% gel and the bands were very smeared. I tried the buffer twice using different voltages (90V and 280V), and the results were pretty much the same.
Any insight as to why this is happening, or what I am doing wrong? Thanks.
Are you using blue dyes in your loading dye mix? These buffers are not compatible with sodium salts and both xylene cyanol and bromphenol blue are sodium salts. I think the recommended dye is cresol orange. I had good success with these gels although I was unable to run the gels at 280 volts. 200 was my maximum.
(i'm a nitpicker) xylene cyanol is a sodium salt, bromphenol blue is not