Thanks for that. I am currently looking for an electrophoresis box. I have everything else I need to do my PCR and run gels, so I will keep this in mind.
I am thinking about getting a gel box from www.iorodeo.com. They supply platinum electrodes and I think it will be cheapest and easiest to get one from them as it should last a while. The tank isn't very big though, which is not great. I should be able to differentiate bands to ~100bp I think though.
Do you have a home lab?
On Sunday, June 24, 2012 6:54:50 PM UTC+10, cory....@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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