Science Fair - HELP NEEDED

[Advait Patil]

Hi,

I am doing a science fair project on how salinity affects the amount of oil degraded by the bacteria Bacillus subtilis.

I have the freeze-dried culture B. subtilis from Carolina scientific, I have rehydrated it and incubated it in a tube in nutrient broth at 30 degrees Celsius for 12 hours, but there is no growth.

Do you think there will be any growth? The rehydration/incubation period for the bacteria is 24-48 hours, and the bacteria are not showing any signs of growth. There are only fifteen days left for my experiment.

If not, does anyone have B. subtilis culture that I can have (not freeze dried; ready to use)? I need to start experimentation asap, so I need culture within the next two-three days. Also, does any place sell B. subtilis locally? This way I won't have to wait for shipping and I can start my experiment. I have nutrient broth, so I only need 2mL, a very small amount. This would be a great help if someone could provide culture.

Thanks!

[Nathan McCorkle]

B.subtilis is a survivor, was your stock culture liquid or dry? One way I've been taught to preserve b.subtilis is with 'potato chip' plates, basically agar with b.subtilis growing that you leave on the lab bench (petri dish closed) until the agar gets so dry it warps and looks like a potato chip! The b.subtilis forms spores and will last years and years and years.

12 hours isn't much time for seeing growth, give it another half-day to get upset! What broth did you use? Did you try streaking some agar plates?

One thing a lot of organisms dont like is being too dilute, so if you used too much broth to begin the first culture, it could be growing super slowly.... Try less volume in the first culture.

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[Nathan McCorkle]

Whoops, I see your stock was dry.

[Koeng]

Contact the bacillus genetic stock center. The professor leading it is an awesome guy and will probably send you some normal Bacillus for free if you ask. 

[Koeng]

BTW i am curious why you'd freeze dry bacillus, they spore well, i mean you can just leave a culture out and they will spore in that

On Saturday, February 22, 2014 10:27:45 AM UTC-8, Advait Patil wrote:

[Nathan McCorkle]

On Sat, Feb 22, 2014 at 3:28 PM, Koeng <koen...@gmail.com> wrote:
> BTW i am curious why you'd freeze dry bacillus, they spore well, i mean you
> can just leave a culture out and they will spore in that

Probably just extra precaution, since it's Carolina they've got the
money to invest in a freeze drier, and if it extends shelf-life it's
likely more than worth it for them as a big company with wide a
variety of stock. So likely they use the freeze drier for the other
strains that require it, and it works for b.subtilis spores too.

The whole going around the line of phase change thing is pretty
awesome! If you're interested Ben Krasnow has made a few DIY machines,
but he's really smart (in a mathematical/engineering sense) and has
tons of tools... so it might take a well-rounded hackerspace or a good
mechanical engineer to pull this off safely. Having one of these
designs in the DIYbio toolbox would be pretty cool! They're required
for getting good resolution with bacteria in electron microscopy, and
can be used to replace VOCs in extractions (i.e. for metabolic
engineering product confirmation).
https://www.youtube.com/watch?v=2wkgBPk0DWg