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maintaining E coli innoculum [View Printable]
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Amritha Nair
Group: Member Posts: 32 Joined: Oct 28, 2007
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I have seen ppl maintain their original E. coli culture in glycerol instead of using plates. Is there a reason why glycerol is used??
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Posted Jul 19, 2008, 0:43 AM |
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samm
Group: Moderators Posts: 409 Joined: Mar 03, 2005
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Usually, large culture collection centers freeze down bacteria such as E. coli in glycerol (different amts usually range from 20-50%), which acts as an anti-freeze, preventing the formation of small spicules of ice that can puncture cells. Storing E. coli in plates in the cold room is a rather inefficient, labor-intensive method, that can lead to strain drift with repeated clonings. If you do want to store them in a cold room/refrigerator (e.g. some transformed cells may lose the vector on thawing), performing stab cultures may actually work out better.
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| Posted Jul 21, 2008, 11:10 AM |
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Amritha Nair
Group: Member Posts: 32 Joined: Oct 28, 2007
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| Posted Jul 21, 2008, 12:45 PM |
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cfish
Group: Moderators Posts: 531 Joined: Sep 21, 2006
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Stab cultures are made by piercing a solid agar medium in a test tube with an inoculating needle covered with the bacterial inoculum. Usually used for cultures of anaerobic bacteria.
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| Posted Jul 22, 2008, 14:31 PM |
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samm
Group: Moderators Posts: 409 Joined: Mar 03, 2005
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| cfish said: | Stab cultures are made by piercing a solid agar medium in a test tube with an inoculating needle covered with the bacterial inoculum. Usually used for cultures of anaerobic bacteria.
| That sums it up perfectly - thanks! Stab cultures are also used for refrigerated storage/culture maintenance, and even RT transport of some strains - however, if you can freeze them down, thats the way to go.
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| Posted Jul 23, 2008, 16:47 PM |
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cfish
Group: Moderators Posts: 531 Joined: Sep 21, 2006
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For long term storage, certainly freeze in glycerol at -80C.
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| Posted Jul 25, 2008, 14:38 PM |
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