Re: [DIYbio] Borax as a superior alternative to Tris

A pain because I have to get it from a scientific supplier instead of the local supermarket.

Also, in Australia there are some pretty strict laws on importing chemicals. It sucks being stuck on an island sometimes. Even to get harmless reagents, I need to get letters from the office for gene technology regulation stating that I am ok to get them. This even applies to Tris. Without that letter most businesses won't even speak to me. One lady even verbally abused me for trying to set up a home genetics lab. She said unless I had a registered business that I had no right to order chemicals from them - even if I wanted to purchase 0.5M NaCl (not that I would anyway). Ridiculous.

So yeah, if I can get it from the supermarket it makes my life a bit easier.

On Tuesday, June 26, 2012 6:38:24 AM UTC+10, Nathan McCorkle wrote:

On Sun, Jun 24, 2012 at 3:53 PM, Cathal Garvey <cathalgarvey@gmail.com> wrote:
> It would be great to use Borax for other buffers, but I gather the borate
> ion is inhibitory to enzymes. Tris is a pain to get hold of, so a nice
> replacement would be really, really cool.

Tris is a pain to get in what sense? The tris /wiki page says/ its
used because its cheap...  (and since everything on the internet is
correct...)    :P

>
>
> On 24 June 2012 14:54, Robert O'Callahan <ropoctl@rice.edu> wrote:
>>
>> I've used SB buffer to run gels. It's nice to be able to use the maximum
>> voltage, but they don't run as well. When loading high concentrations of DNA
>> the migration distance can do weird things, but it works well for
>> preparatory gels when you need good separation quicker.
>>
>> -Rob
>>
>>
>> On Sun, Jun 24, 2012 at 3:54 AM, Cory Tobin <cory.tobin@gmail.com> wrote:
>>>
>>> > I read a paper a while ago showing a comparison of
>>> > different buffers, and feel like the borax ones weren't
>>> > as good for certain fragment sizes or something...
>>> > what I mean is, I feel like there was some tradeoff
>>> > between TAE and borax buffers
>>>
>>> SB doesn't do well with high molecular weight fragments, like >8k or
>>> so.  TAE works better in that case.
>>>
>>>
>>>
>>> > But this could possibly made even better by reducing the
>>> > Na in the solution.
>>>
>>> True.  Lithium is better but it's expensive.
>>>
>>>
>>>
>>> > Also, the above papers said that you don't really need
>>> > EDTA either. The use of EDTA is kind of a left-over
>>> > from RNA gels that persists in DNA gels for some reason.
>>>
>>> This is typical of biology at universities.  Someone finds a protocol
>>> that works and keeps using it without determining what parameters are
>>> actually important.  There is no incentive to try running gels with
>>> missing components because discovering that EDTA is not needed will
>>> not get your NIH grant renewed or help you get tenure.  From any
>>> individual's perspective it's better to just continue using protocols
>>> that work and focus on their research, especially considering that gel
>>> buffers are a very minor portion of any research budget.  Although,
>>> this might not be the case for diybio.
>>>
>>>
>>>
>>> > Does anyone here work with Borax? It is readily
>>> > available from the supermarket and I would be
>>> >keen to hear of anyone else's experiences
>>>
>>> Yeah, it works well.  I've used both methods: (1) Borax only and (2)
>>> NaOH + Boric acid.  Both work fine.  One thing to note is that you
>>> can't make a 50X stock of SB like you can with TAE.  The max I go is
>>> 20X and even then I still get some precipitation.  It's a bit
>>> inconvenient but not enough to make me stop using it.  Also, if you
>>> plan on running your gel at 300V for more than 10 minutes, make sure
>>> you have a box with large reservoirs or you'll melt the gel.
>>>
>>>
>>> -cory
>>>
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>>>
>>
>>
>>
>> --
>> Rob O'Callahan
>> M.S. Civil & Environmental Engineering 2012
>> George R. Brown School of Engineering | Rice University
>> E: rpo1@rice.edu<mailto:rpo1@rice.edu>| P: 847.641.1285
>>
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>
>
>
>
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--
Nathan McCorkle
Rochester Institute of Technology
College of Science, Biotechnology/Bioinformatics

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