Inoculating bacteria

About microscopic forms of life, including Bacteria, Archea, protozoans, algae and fungi. Topics relating to viruses, viroids and prions also belong here.

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NiceOnTheIce
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Inoculating bacteria

Post by NiceOnTheIce » Thu Oct 14, 2010 12:26 am

Hi there,
I was hoping someone could tell me why you should only use 1 bacterial colony to inoculate a tube of broth, vs using many different colonies. I know that using many colonies increases your risk of contaminating your culture, but what exactly does that mean?
Thanks.

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canalon
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Post by canalon » Thu Oct 14, 2010 1:01 am

that if you pick one typical colony you are likely to pick a good one, but if you scrape a plate where some contaminant might be lurking, you might miss it in the crowd.
Patrick

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any proof. (Ashley Montague)

NiceOnTheIce
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Re: Inoculating bacteria

Post by NiceOnTheIce » Thu Oct 14, 2010 1:19 am

That makes sense.
But continuing on from that point: Does each colony of bacteria differ from one another? Could that be another way that the culture would become "contaminated" by using 2 or 3 colonies? Or are the colonies pretty much identical?
Thanks.

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JackBean
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Post by JackBean » Thu Oct 14, 2010 6:12 am

let say, you have transformed your bacteria with some plasmid. In such a case, the amplification and ligation did not have to work well and really each colony can have sligthly different version, with some mutations. If you pick only one colony, it probably comes from only one original cell and should be homogenous, but can differ from other colonies on the plate
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

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canalon
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Post by canalon » Thu Oct 14, 2010 8:52 pm

Jack is true.
But from a frozen stock (or even one agar) if everything has been done correctly, in all likeliness all colonies are equivalent (there might be mutants, but that happen also within the colony, so it is not very relevant in general).
So you can use more than one colony if the experiment requires it. Just take multiple isolated colonies, to make sure that you are not picking something else at the same time. For example if you try to isolate E. coli from a soil containing some Proteus sp. multiple repeated steps of picking isolated colonies on fresh plate will be necessary to get rid of the contaminants.
Patrick

Science has proof without any certainty. Creationists have certainty without
any proof. (Ashley Montague)

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Post by alchilito » Sat Oct 23, 2010 12:45 am

Theoretically, every colony is clonal, stemming from single isolated cell that grew up to being visible macroscopically. Jack's post are correct.

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