Re: [DIYbio] Re: Promoter Testing

On Fri, May 15, 2015 at 10:09 AM, Linden <ff.lindenberg@gmail.com> wrote:
> Hey guys,
>
> Thanks Nathan and Sebastian, I will compare the promoters. Just to clear it
> out, there is no way to know the amount of GFP inside the cell using only
> the spectrophotometer results? Lambert-beer equation doesn't apply in this
> case?

Beer-lambert does still apply, but measuring cell solution is going to
give a bit of a different result than measuring purified GFP. There
are going to be a lot of other molecules in the cytoplasm, in the
growth media, etc... the cell membrane will be in the way. So as long
as you compare your results to previously published (or your own
side-by-side) values that followed the same methodology (measuring
cell solution) you will get a meaningful answer. My guess is that even
something like cracking the cells open with a drop of soap or boiling
lysis is going to change the signal measured. Being consistent is key.

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