Buying bacterial reagents from local drug store

[izaak coleman]

Hello DiyBioists,

This is my first (hopefully of many!) contact with the DIYbio community! I plan to drag myself and my lab to a backspace soon, instead of lazing around on the weekends as per!

So, I am currently trying to grow around 10 weird'n'wonderful bacterial species, with the aim of imaging the rotary motor at the base of their flagella using Electron Microscopy. However, I have run into a jam when it comes to reagents. Two of my species' media require a 'vitamin solution' in a very small quantity, and it turns out that this vitamin solution is effectively a concoction of common B vitamins...

To buy these vitamins from sigma and other pro-chem vendors, each is roughy £30-100. This is a hefty sum when I need around 10 vitamins in total to produce the vitamin solution! FYI, this high price is due to the assurance of >99% purity.

However, I have found all the vitamins I require in my local drug store, and the grand total for all the reagents is £30! So my question is, has anyone used vitamins (or any tablet form reagents bought from a drug store) to grow bacteria? Any advice? From what I can tell, the bulking and stabilising agents are all harmless to bacteria... but if anyone knows anything to avoid that would be helpful!

List of bulking agents:

Microcrystaline Cellulose

Dicalcium Phosphate

stearic Acid

Silicon Dioxide

Magnesium Stearate

Cheers!

[Jeff Backstrom]

Do you have access to a centrifuge, even a blood centrifuge?

Dissolve your vitamin tablets in a bit of distilled water. Centrifuge. The B vitamins are soluble, while many of your excipients are not. You will greatly reduce the non-vitamin components of the tablets by using a pipettor to withdraw some of the solution; there may be some material on top (I would guess the stearic acid is hydrophobic and will float anyway, for example), so withdraw from between what floats on top, and what insoluble material there is on the bottom.

An option would be to purchase your medium (add-water-and-autoclave style) from a commercial supplier, vitamins and all. Perhaps not as much DIY bio as would be nice, but if it saves you that much money, maybe it is for the best.

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[Dakota Hamill]

What species are you specifically trying to grow?  That could help to find alternatives or specialized media recipes.   If you have access to an electron microscope, don't you have access to the bio-departments stock of media reagents?  

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[Nathan McCorkle]

Are you going to try to image live and active cells? In an
environmental SEM/TEM? Could you provide some article references if
so?

I don't remember how the videos I've seen of flagella moving were
recorded, I assumed it was fluorescense or some other optical
technique (quick google shows fluoro and dark-field, but not eSEM).

On Sun, Aug 10, 2014 at 11:54 AM, izaak coleman <izaakm...@gmail.com> wrote:

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