Sodium Boric Acid as TAE replacement - Does anyone use SBA? (Sep/09/2005 )

Hi,

I run a million gels a day and want to switch over to using SBA instead of TAE or TBE because it allows for faster electrophoresis. I've tried numerous recipes for this buffer, but my bands are always distorted. If anyone uses this buffer and has positive experience with it, could you please post the exact recipe you use to make it? Basically I'm following the protocol outlined by Brody and Kern in:

"Sodium boric acid: a Tris-free, cooler conductive medium for DNA electrophoresis."
Johns Hopkins University School of Medicine, Baltimore, MD, USA.
PMID: 14989083

Thanks,
Hank

sodium boric acid 10 mM final
EDTA 1 mM
ethidium bromide 1 mg/l

i usually prepare a 10x solution from sodium borate salt

no problem with agarose

Seb_

Hi Seb, thanks for the reply.

I noticed that the paper also mentioned that you could use boric acid and then pH it with NaOH. Have you ever tried this? This is what I've been attempting since I don't have sodium boric acid readily available.

Thanks,
Hank

i remember having some troubles after a mistake, taking boric acid instead of sodium borate, but at this time i have not checked the pH (but EDTA was at pH9.0)...

NB: for sodium boric acid, i mean Na2B407, also called Borax http://en.wikipedia.org/wiki/Borax

Seb_

That's it!!! I made the exact same mistake!

Thanks!
Hank

Edit: What a lame way to make my 100th post

Hi,
TAE and TBE are truly ancient and poor buffers. SB should be fine but LB(lithium boric acid) is truly the king of all buffers for excellent resolution. CHeck out:BioTechniques® October 2004
Volume 37, Number 4: pp 598-602
Short Technical Reports
Ultra-fast high-resolution agarose electrophoresis of DNA and RNA using low-molarity conductive media

This paper came out after the one you mention. Also, there is a great history lesson in Analytical Biochemistry about these buffers. Please also refer to the technical advice at www.fasterbettermedia.com where you can buy these buffers for excellent and fast resolution. Lithium acetate is the appropriate switch for upper molecular weights (over 8kb or so).

Please feel free to contact me if you have any problems--our whole Lab @ Hopkins has switched over and have saved tons of money and time!

I'm still a little confused as to the right substance for this buffer -- is this what I want to use?

yes

Thanks!

Hi,

I read the LB paper from 2004 as well and found it very interesting. Since the components for SB are readily available to me, however, I'll most likely try that out first. Have you ever experienced problems using fragments purified from SB or LB gels? I'm assuming there shouldn't be a problem, but just curious. Also, have you basically stopped on LB, or are you investigating the potential for other buffers as well?

Keep up the good work!

Thanks,
Hank

Hi,
TAE and TBE are truly ancient and poor buffers. SB should be fine but LB(lithium boric acid) is truly the king of all buffers for excellent resolution. CHeck out:BioTechniques® October 2004
Volume 37, Number 4: pp 598-602
Short Technical Reports
Ultra-fast high-resolution agarose electrophoresis of DNA and RNA using low-molarity conductive media

This paper came out after the one you mention. Also, there is a great history lesson in Analytical Biochemistry about these buffers. Please also refer to the technical advice at www.fasterbettermedia.com where you can buy these buffers for excellent and fast resolution. Lithium acetate is the appropriate switch for upper molecular weights (over 8kb or so).

Please feel free to contact me if you have any problems--our whole Lab @ Hopkins has switched over and have saved tons of money and time!