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Grow multiple Ecoli colonies in a single liquid media tube

Ecoli Colony LB media liquid

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#1 SamSam

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Posted 30 May 2013 - 08:27 AM

Hi,

I have a plated library of random mutations in my gene. My question is, can I , theoretically pick all the colonies into a single liquid media culture and allow them to grow for a few hours and then plate the culture onto a plate?
I do not have the ligation mix for the library to do another transformation so I am hoping I could just reuse these colonies so to speak.
I would obviously include the appropriate anitbiotics in the liquid media.

Thank you.

#2 Trof

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Posted 30 May 2013 - 09:26 AM

You have library on a plate.
You want to grow inoculates for a few hours (in which they won't grow much) and then put them on a plate again...

So you start with a plate and end with a plate. I can't really see what exactly are you expecting from that, apart from the chance, that some of the clones would be lost in dilution.

Our country has a serious deficiency in lighthouses. I assume the main reason is that we have no sea.

I never trust anything that can't be doubted.

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#3 SamSam

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Posted 30 May 2013 - 03:43 PM

Thanks, Trof,

I should have mentioned that I want to plate the library on a new media with some specific additives.
The cloned gene also codes for an enzyme which is secreted so I want to check its activity via cleavage of a product spread on the new plates.

#4 Trof

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Posted 31 May 2013 - 12:00 PM

Will your bacteria grow right away on your new media?

If they will, from my point of view, the easiest way would be to do a replica plate, as mentioned for example in this article.

Our country has a serious deficiency in lighthouses. I assume the main reason is that we have no sea.

I never trust anything that can't be doubted.

'Normal' is a dryer setting. - Elizabeth Moon


#5 notjustalabtech

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Posted 09 June 2013 - 10:26 AM

You could, but how would you know which colony was which? And what if for whatever reason, some colonies grow faster than others so you might lose the slower ones. It's better to do it from plate to plate. This wouldn't work unless the colonies were cultured one by one in separate tubes. You would waste a lot of time in trying to identify your colonies and it might make people skeptical of your results. You could try streaking a plate in sections, like the one in this picture: http://www.tcnj.edu/.../RNAiStreak.jpg





Also tagged with one or more of these keywords: Ecoli, Colony, LB media, liquid

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