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Specialty Chemistry Forums => Biochemistry and Chemical Biology Forum => Topic started by: Henry246 on January 19, 2015, 01:19:33 AM

Title: Bacteria sample generation help
Post by: Henry246 on January 19, 2015, 01:19:33 AM
Hi guys,

I'm currently trying to perform some soil tests involving cyanobacteria contamination, and I need a way to create my own cyanobacteria samples. I plan to grow some bacteria cultures in a petri dish but I am unsure of how to get the bacteria back out and into a liquid sample after it has grown. Any advice you can provide on the topic is much appreciated.

Just a quick bit about myself fyi, I am studying to be a civil engineer and have very little chemistry background, and have found myself a little out of my depth when working on a very chemistry-based project, enjoying it though!
Title: Re: Bacteria sample generation help
Post by: Arkcon on January 19, 2015, 08:44:43 AM
Hmmm... this is, on a level, basic microbiology.  If you've grown a colony of bacteria on an agar plate, you use a sterile loop, to transfer some of the colony aseptically to a tube of broth.  Here's a cartoon based basic tutorial:  http://www.biotopics.co.uk/microbes/tech1.html

I don't know however, if what I'd done as an undergrad to general microbes applies to cyanobacteria, which need different nutrients, and sunlight to grow.  You may need to talk to a phycologist http://en.wikipedia.org/wiki/Phycology
Title: Re: Bacteria sample generation help
Post by: Henry246 on January 20, 2015, 02:38:26 AM
Thanks for that, I hadn't read of the liquid broth technique before. My issue now is that sealing the broth bottles will create anaerobic conditions and prevent the cyanobacteria from growing, however leaving the lid off could result in contamination of the culture.
Title: Re: Bacteria sample generation help
Post by: Arkcon on January 20, 2015, 05:31:56 AM
I'm finding out that cyanobateria are very different to culture, compared to other bacterial culture methods.  About all they have in common is the name "bacteria", and that's just historical.  Try this reference for starters. http://aem.asm.org/content/57/5/1448.full.pdf
Title: Re: Bacteria sample generation help
Post by: Babcock_Hall on January 20, 2015, 05:45:46 PM
It is possible to use foam plugs or cotton balls wrapped in cheesecloth to allow for gas exchange under sterile conditions.
Title: Re: Bacteria sample generation help
Post by: Yggdrasil on January 20, 2015, 08:21:36 PM
Thanks for that, I hadn't read of the liquid broth technique before. My issue now is that sealing the broth bottles will create anaerobic conditions and prevent the cyanobacteria from growing, however leaving the lid off could result in contamination of the culture.

You can buy cell culture tubes that have a cap that goes on loosely, allowing for gas exchange, but still covering the culture to prevent contamination.
Title: Re: Bacteria sample generation help
Post by: Henry246 on February 01, 2015, 01:34:34 AM
Hey Arkcon can I ask how you managed to find that paper specifically on cyanobacteria culturing? I think my research techniques could use developing, given that I spent hours and hours looking for pretty much exactly that and was unable to find it.
Title: Re: Bacteria sample generation help
Post by: Arkcon on February 01, 2015, 11:17:01 AM
I just Googled for it and picked the first, simplest one.  Like I said, this is really a specialized task.
Title: Re: Bacteria sample generation help
Post by: Babcock_Hall on February 02, 2015, 08:46:38 AM
If you don't already have a good microbiology laboratory textbook, then obtaining one might be a good idea.