Re: [DIYbio] gel electrophoresis vs liquid chromatography?

I just did HPLC today on caffeine and it was pretty easy... I was
checking column prices last week on ebay and they seemed like around
$50 for used... but what about new?

What are the ideal DNA columns? Is there an easy/cheaper DIY
compromise/substitute available? (maybe silica?)

On Wed, Feb 18, 2015 at 11:57 AM, GO <gobiowork@gmail.com> wrote:
> We can also co-pass a already known strands with some fluoro or other
> label that can be detected at wavelength other than 260 nm (this would serve
> as a ladder in electrophoresis).

Concern might be that the dye dislocates during chromatography...

> Also its more difficult to collect DNA
> later in gel compared to HPLC. Liquid chromatography can be used to detect
> other things as well (not just charged DNA) so it's even more useful for DIY
> people.

I didn't collect elutions, but I think I would have just had to figure
out the lag time between the detector and the end of the waste tube.
It certainly would have been easy. The one problem I could see is that
while the sample loop was only 100uL, it was recommended to flush with
1000uL (to clean out any trace of previous sample) and I was using
more like 2 or 3 mL since we had plenty. So for people with only 50uL
or even 500uL, we need to figure something out... as a gel can be
loaded with something like 10-20uL. I am not sure if diluting it to
1000uL would be smart or not... I am sure the required volume could be
decreased, but there is still the flush volume to prevent
cross-contamination... I guess this could be switched from sample
being used to wash/rinse to DNA buffer.

I think the high-pressure generation and injector valve is the hard
part of this. Maybe a good reason to go after capillary (or capillary
gel electrophoresis) electrophoresis instead... no pressure to worry
about, so adding sample shouldn't be much different for end-users used
to loading gel wells.

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