Re: [DIYbio] Re: My first successful CRISPR experiment on E. Coli ?

On Fri, Dec 23, 2016 at 10:22 AM, Hugues <laliberte1965@gmail.com> wrote:
> Hi,
>
> the normal Agar LB and the Strep LB came in powder form in 2 tubes with the
> kit. I just followed the instructions and dissolved the powder in water,
> then heated in microwave a bit until it became clear,

As mentioned in this comment, degradation might be an issue above 75
degrees F (it says prior to some patent invention, but from reading
the patent it seems like they just found a good pH and buffer system
to help get around this issue):
https://groups.google.com/d/msg/diybio/GsPMAfyfAkU/R2NmH4XZad8J

So my point is, maybe you overcooked the streptomycin... you want to
be very gentle on the solution as you microwave it (pulses of 10 to 30
seconds at a time depending on volume of liquid, then swirl to mix,
then heat again, swirl, heat)... this is even more important if your
microwave lacks a rotating plate (you could get hotspots that are
super-heated, and thus more easily degrade unstable molecules in that
volume of liquid).

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