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

The odd thing about what you said is "For me, these whitish streaks do
not seem to be growing colonies, the streak appeared rather rapidly
after i streaked both strains, once they were dried, but then they
stayed like this."

So if you followed the instructions, you'd have dropped 100uL onto the
plate, then spread it around. From my memory, and the last time I
spread cells was probably 4 years ago, 100uL is reasonably wet... so I
would not recommend mixing strains on the same plate/dish.

Continuing, if the appearance of the agar hasn't changed ("but then
they stayed like this"), seems like you might have over-dried to begin
with. A spread of culture, assuming you don't scrape/scratch the agar,
should look like a dirty/oily window, rather than a window with
mucus/paste obviously spread over it (what I'd call yours).

A question I have, aside from why you tried to plate so much of two
cultures (100uL each) onto a single plate, is, did you mix the culture
before plating? I don't think this really matters experimentally, but
just thinking about reasons why your dry streaks look so thick (i.e.
maybe you didn't mix, sucked up the cell-pellet that settled at the
bottom of the tube).


On Thu, Dec 22, 2016 at 12:30 PM, Hugues <laliberte1965@gmail.com> wrote:
> They were 15h old at the moment of the picture.
>
> What makes you think it did not work ? I did not have any dots at the beginning, now there are, so something is growing.
>
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--
-Nathan

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