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maintaining E coli innoculum

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Amritha Nair

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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
samm

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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
Amritha Nair

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what are stab cultures?

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Posted Jul 21, 2008, 12:45 PM
cfish

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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
cfish

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Here is a link for a basic procedure for the preparation of slant and stab cultures.

Link: http://cfcc.edu/faculty/jjenkins/courses/msc180/lectures/slant_stab_broth.doc

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Posted Jul 22, 2008, 14:34 PM
samm

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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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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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