TBE Buffer? - (Jun/08/2010 )
Anyone know how concentrated I can make TBE buffer - I've seen up to a 10x recipe, but I'd like to make 50x - just not sure that all that boric acid will go into solution.
Precipitation of TBE frequently occurs, especially when a 10x stock is prepared. (Molecular Cloning, Sambrook et al) 5x stocks are more stable. Autoclaving will retard this reaction. Alternatively, filtration shortly after preparation through a 0.2-0.45 µm cellulose acetate or cellulose nitrate membrane will reportedly allow a 20x stock to be prepared. (Krainer, Adrian R.; Mayeda, Akila. 1991. Long-term Storage of Concentrated Tris-borate-EDTA Electrophoresis Buffers without Precipitation. Benchmark, Biotechniques, Vol.10, No.2, p.182 ) 30x stocks prepared by this method are stable for 2 weeks. Filtration suggests that nucleation of salt crystals by dust particles or other microscopic insoluble material is responsible for the precipitation problem. If turbidity develops in filtered buffer stocks, autoclaving for 5 minutes at 110°C will remove cloudiness. 10x stocks that have massive amounts of crystallization can still be used to prepare 1x TBE without affecting the resulting voltage, watts or amps.
IMHO, I will not prepare anything more than 5x for TBE buffer. I used to prepare a lot of 5x buffer but in the end all was crystallized. Since I use only 0.5x buffer, I only prepare 1x and this last me for quite long as well.
Just curious: why you want to do up to 50x ? Do you think after dilutions into 1x or 0.5x , the pH and buffer strength will be less likely to be affected (unless yours is neutral water)?
For 1 Liters of 10x TBE, you need: 108g Tris base, 55g Boric acid, 40mls 0.5M EDTA (pH 8.0)
(I read somewhere in this forum says that prepare 1M EDTA is impossible)
Since you want 50X TBE, I am skeptic if you can really dissolve 540g tris base, 275g Boric Acid, 200ml 0.5M EDTA into a 1 liter solution. Adding 815g powder of tris base and boric acid takes almost a full bottle, and adding 200ml EDTA I'm skeptic about how will it dissolve the tris base and boric acid. which the salt amount is more than the solution. Furthermore, I think you hardly top up dH20 into your bottle.
So,in my humble opinion, to prepare 50x TBE is impossible. If you can do it please let me know how.
Anyway, I was thinking if I prepare 1 liter 50x TBE and I got 100 liter to use... not a bad idea..
We prepare several litres of 10x TBE and then autoclave it and we never had problems with crystalization.
Can't speak to the logistics of making it on the bench yourself, but a colleague used to use tablets he purchased somewhere that would dissolve to prepare 50x TBE. He seemed to like them, but I can't remember what vendor they were from.