How do you grow specific types of bacteria?

[V Shenoy]

Hi I am new to Diy bio, and I was wondering if I could get some help from you guys. I was wondering if any of you had any advice on trying to grow specific types of bacteria. By this I mean that I want to grow bacteria with specific traits. I am trying to do this on a fairly low budget (so no buying bacteria) but I have sufficient lab space, a microscope, and other equipment needed for growing bacteria. I would really appreciate it if you could help me. Thanks! 

[Nathan McCorkle]

look up selective growth media .

On Dec 2, 2013 5:44 PM, "V Shenoy" <vshe...@gmail.com> wrote:

Hi I am new to Diy bio, and I was wondering if I could get some help from you guys. I was wondering if any of you had any advice on trying to grow specific types of bacteria. By this I mean that I want to grow bacteria with specific traits. I am trying to do this on a fairly low budget (so no buying bacteria) but I have sufficient lab space, a microscope, and other equipment needed for growing bacteria. I would really appreciate it if you could help me. Thanks! 

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[Koeng]

Buy the strains? Then seed cultures

[Tom Hodder]

On 2 December 2013 01:58, V Shenoy <vshe...@gmail.com> wrote:

Hi I am new to Diy bio, and I was wondering if I could get some help from you guys. I was wondering if any of you had any advice on trying to grow specific types of bacteria. By this I mean that I want to grow bacteria with specific traits. I am trying to do this on a fairly low budget (so no buying bacteria) but I have sufficient lab space, a microscope, and other equipment needed for growing bacteria. I would really appreciate it if you could help me. Thanks! 

Hi,

I am probably not that far ahead, because I only got a microscope a few months ago, but I found that once I could make up agar plates and inoculate with samples confidently, I was able to take swabs off all sorts of stuff and inspect it under the microscope.

Once I had worked out how to get the bacteria to spread out nice and evenly on the agar, it was possible to identify them as cocci, rods, staphylocci, dicocci, etc just under the 40x objective/20x ocular magnification.

After that I started to learn about gram staining, and gram positive and negative, and now I am looking at using growth medium and ampicillin to select for particular strains.

However if I as starting again, this time I'd get the basic handling of bacteria in a sterile fashion as basic skill first, and then move on to selecting for traits when you are confident.

Cheers,
Tom

 

[Cathal Garvey]

Depends what you want to isolate. Some are very easy to isolate because
they have traits (ability to grow at low pH, high salt, low
temperature, without sugar, with odd sugars, with alcohol... etc) that
can be easily selected for. Others are hard to isolate but easy to
identify. I would say that most are easy to grow but hard to identify,
and most are medium difficulty to isolate by differential methods
without specialised ingredients.

However, some species will be next to impossible to identify or isolate
without special equipment or ingredients, often species you would
expect to be easy. I spent a good few evenings digging into Bacilli,
trying to find a way to isolate *just* bacillus subtilis, or even its
close relatives, from soil samples, and not B.anthracis or B.Cereus.
Sadly, it turned out to be the sort of thing that requires equipment,
technique and sometimes ingredients that won't be on-hand for most. I
could do it, because I'm a microbiologist, but it wasn't worth trying
to write up a how-to.

On the other hand, isolating V.phosphoreum is *easy*: You leave an
unwashed, unfrozen squid in a fridge (one you're not using for food!)
in a salt-water + glycerol solution until it goes bad, and look at it
in a dark room for glowing spots. Isolate the blue ones, grow on LB
agar + glycerol + salt to 32g/L, at 4C in the dark. Very little else
will grow at 4C at that much salt, except perhaps S.aureus, and even
then competitors will grow very slowly, and *don't glow in the dark*.

So, it depends on the species. Some are easy, some are hard. The main
thing is knowing!

[Koeng]

What concentration of salt water/gycerol do you use initially?

[Avery louie]

http://tequals0.wordpress.com/2011/09/29/isolation-of-vibrio-fischeri-from-seafish/

information about isolating vibrio can be found here.

--A

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[Patrik D'haeseleer]

Also, extremophiles can be quite easy to isolate, for the simple fact that almost nothing else will grow at the conditions they like. For example, you can actually isolate halophilic (i.e. salt-loving) archaea from a wide range of gourmet salts, simply by using saturated salt growth media (and plenty of patience).

Patrik

On Monday, December 2, 2013 5:28:01 PM UTC-8, Cathal Garvey wrote:

[Mega [Andreas Stuermer]]

I didn't add glycerol to it, and also got glowing from sea fish.

Btw., you could add myristic acid which may enhance the glow.


Which brings me toa new idea, supplementing glowing E. Coli with the fatty acid, to get pVIB on steroids. :D

[Koeng]

You could also optomize the RBSs of all the pVIB proteins, https://salis.psu.edu/software/. The RBS will promote over expression of the pVIB proteins, even though that might screw over your cells because of such high expression

And also get better promoters, you can PCR them from the genome... all you gotta do is boil a tube of E coli and use it for PCR xD

[Mega [Andreas Stuermer]]

Sounds nice. Does that optimization tool also work for gram positives? I mean secondary structure avoiding yes, but consensus sequence?

After all as with each diy project... If you engineer it, then engineer it to work in all bacteria :D



Btw, I would recommend sea fish too. Easy to start with, and nice glowing effects. Just make sure not to breed V. Chloerae (4°C)

[Koeng]

you can optimize to any bacteria... however I think that its a bit broad to make it try to work in all bacterium, i mean you most likely are not gonna use it in anything but e coli (maybe bacillus)

[Andreas Stuermer]

Yeh, Bacillus Subtilis is nice. GRAS and sporulating (so easy to keep for years/decades dried)

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