low molarity borate and acetate agarose gels
What are people's experiences using the Brody and Kern (Biotechniques
36:214 and 37:598) low molarity sodium borate and lithium acetate
agarose gels? They do run very fast with little heat, but we often
get very smeared bands, with borate performing better than acetate.
There is a lot of band quality variability from run to run. Any tips
would be appreciated!
Re: low molarity borate and acetate agarose gels
Buffers for fast agarose gel electrophoresis
Sodium Borate Buffer – 10 mM sodium boric acid (1X working solution is either 5 mM disodium borate decahydrate or 10 mM NaOH adjusted to pH 8.5 with boric acid, e.g, 8g NaOH, 47g boric acid in 1 L water to make 20X buffer, pH ~ 8.0).
Lithium Acetate Buffer – 5 mM lithium acetate, for separating DNA fragments >3 kb. (restriction digestion).
Lithium Borate Buffer – 1 mM lithium boric acid (lithium hydroxide pH buffered to pH ~8.5 with boric acid). For small DNA fragments (PCR products). Lithium Borate Buffer has better resolution than Sodium Borate Buffer for small DNA fragments.
Operative run gels between 200 and 300 Volt
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