Re: [DIYbio] Isolation and sequencing of RNA

On Thu, Mar 19, 2015 at 1:51 PM, Luke <lukas.lukasmoser@gmail.com> wrote:

> So if you let the Thermocycler copy the DNA 54 times and the PH-Meter can
> notice the difference of 0.1 PH, it works?

Some things that immediately come to mind relate to the ionic strength
of the solution. Also, you'd need to measure the pH before adding free
nucleotides, as these will contribute as acids I think. Measuring
low-concentration DNA solution, then adding master mix and amplifying,
then purifying seems like opportunity for lots of transfer loss to
affect the results. Not to mention how much the ion-selective
electrodes (ISE) tend to drift... you'd also need a way to ensure the
ISE didn't cross-contaminate your samples unless you weren't
particularly interested in using them.

I wonder how you would do with a visual pH indicator, but at that
point you're just trading the use of something like 260nm light for
something higher in the visible (and it's less direct indication).

In theory it sounds like a reasonable idea to try out... but I would
recommend doing a good prior-art literature search as it does seem
like something someone could have tried before.

Should you try this, let us know the results of course!!!

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