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well, shit.
Like pretty much everyone, I've been using Tris-based buffers for nucleic acid electrophoresis since I started doing it. Turns out that the buffering capacity of the solution makes no real difference, and what you really want is a solution that doesn't carry so much current (and therefore doesn't generate as much heat; I've melted TAE/agarose gels before). I guess something was lost in translation between the interview and the article, because "carries a voltage" is meaningless to me. Also, I note that Kern and Brody have "filed for a provisional patent on the sodium boric acid solution" -- bwahahahaha! Good luck enforcing that. (In defence of the researchers, I suspect that beancounting shitbags at Johns Hopkins have made such idiocy mandatory.) What's great about this is that everyone has been doing it the same way for thirty years, not bothering to think about improving the method since it worked well enough and, you know, that's the way everyone does it. Now these guys come along and deliver a smack-your-forehead moment to every molecular biologist in the world. *smacks self in forehead* What's bad about this is that my one burning scientific ambition is to get a methods paper like this published, and I'm insanely envious. Why didn't I think of this? (Don't answer that.) Comments Ok, this is cool stuff, but what is sodium boric acid? Do they mean sodium tetraborate decahydrate? Dr Dietz: yes, they do. From the paper that prompted this entry (Biotechniques. 2004 Feb;36(2):214-6), the relevant section is: 1× SB consisted of 5 mM disodium borate decahydrate or 10 mM sodium hydroxide, pH adjusted to 8.5 with boric acid. A 20× stock solution of pH 8.0 produces the appropriate pH upon dilution to working strength; stock solutions of 10−50× can also be prepared and stored without difficulty.In February, I made up a 20X solution using the NaOH method and have been using it with good success since. I cannot achieve quite the speed the authors describe since I still get some heat buildup, but I could probably do better if I made the solution up very carefully and optimised for the gel rig I use. I routinely run 0.7% agarose gels (8cm x 6cm x 0.5cm) at 70-100V using this buffer and get very good separation. Perhaps of further interest: Kern and Brody make two other papers available via their company website. The earlier Biotechniques paper is no longer available there, but I have a pdf of it if anyone wants it. senn, we recently ordered a bottle of the SBA solution from Drs. Brody and Kern. It's great stuff. Your're right about it running a bit hot at the voltages they recommend -- we diluted it to .5X and it's nice and cool. Post a comment |
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Great piece. Good choice of subject. You obviously get it.
Check out Cancer Biology & Therapy 2004, the Sept 11 issue.
"Stagnation and Herd Mentality in the Biomedical Sciences."