Re: [DIYbio] 200$ Real-Time PCR Thermocycler

On Tue, Jul 4, 2017 at 2:13 PM, Gabriele Borelli <borelli.g92@gmail.com> wrote:
> - Not quantitatively accurate as the top-notch Roche thermocycler, but more
> than enough precise to reliably discriminate between "DNA has been
> amplified" and "DNA has NOT been amplified"

This will scare off the more professional/power users... experiments
are hard and expensive and maybe use rare DNA, these folks don't want
to waste their time, effort, and precious samples if the hardware has
not rigourously been tested, calibrated, normalized, etc. You'll need
to do environmental stress testing (test in hot environment, cold
environment, for long periods of time in both... and prove that in all
cases, your performance quality remains high and consistent). Quality
Control is king.

> - SYBR green is not necessary.

Why isn't it needed? Do you use UV detection? If so, what dosage over
the whole run (I guess watt-hours, or something with lux, etc)... this
could have implications on the fidelity of the DNA (UV breaks bonds).

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