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Growing E. coli in 96 well?

Growing E. coli in 96 well? - Protocols and Methods Forum

Growing E. coli in 96 well? - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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  #1  
Old 01-06-2004, 05:01 AM
Yoram Gerchman
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Default Growing E. coli in 96 well?



Greetings netters, anyone had good experience
growing E. coli in 96 well plate? What plates and
what cover did you use?
Thanks Yoram

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  #2  
Old 01-06-2004, 05:59 AM
EK
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Default Growing E. coli in 96 well?

Depending on your application, your options could very from conventional
flat or round bottom (~300ul per well) 96-well plates to so called deep-well
plates that can be up to 2.5 ml per well. You can even use 96-well PCR
plates from ABI, but those give no much advantage in terms of volume (same
300ul/well), though might be more convemient (conical chape) if you need to
spin down cells afterwords. Plates can be covered by a sticky foil, special
air transparent film, or multiwell stopper sheets. In my experience, 200ul
headspace on top of 100ul of culture provide enough aeration to sustain an
overnight growth, which I believe in fact consists of a respiration step
followed by semianaerobic fermentation step in fast-growing cultures. If
aeration is important, I would use deep-well plates with minimal culture
volumes per well. Sorry for not providing specific information about the
plates and covers. Everything can be found in e.g. Fisher or VWR catalogue,
and then, if necessary, in original manufacturer's catalogs (e.g., Costar,
Denville, etc.) if items of specific need are not listed in those general
retail catalogs.
Emir.

"Yoram Gerchman" <[Only registered users see links. ]> wrote in message
news:[Only registered users see links. ]...


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  #3  
Old 01-06-2004, 01:43 PM
Tom Knight
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Default Growing E. coli in 96 well?

To add to EK's comments, you might want to look at the Eppendorf
benchtop heater/shaker with the 96 well plate attachment. It has a
short-throw shaker, which is really important for small volume plates
and is quite convenient.

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  #4  
Old 01-06-2004, 03:38 PM
Louis Hom
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Default Growing E. coli in 96 well?


I bought my gas-permeable plate covers from research products
international.
--
__________________________________________________ ____________________________
Lou Hom >K'93
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[Only registered users see links. ]
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  #5  
Old 01-06-2004, 09:18 PM
Walter Schick - BioSepCo
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Default Growing E. coli in 96 well?

Further add to comments is a neat two plate shaker from ATR Biotech in the
US--we have used it in a cold room and run from 10deg C to 55 degrees.

The microtec is designed just for microplates--no attachment needed. Here
is the web page:

[Only registered users see links. ]

Walt Schick
BioSepCo US


----- Original Message -----
From: "Tom Knight" <[Only registered users see links. ].mit.edu>
To: <[Only registered users see links. ].ac.uk>
Sent: Tuesday, January 06, 2004 5:43 AM
Subject: Re: Growing E. coli in 96 well?




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  #6  
Old 01-13-2004, 03:17 AM
Eric R. Hugo
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Default Growing E. coli in 96 well?

Tom Knight <[Only registered users see links. ].mit.edu> wrote in
news:[Only registered users see links. ].mit.edu:


I seem to remember an article where potassium nitrate was included in the
medium as an electron recipient thus allowing the E. coli to grow fairly
robustly anaerobically (although slower eg. 48h instead of overnight). As
long as you are not in a hurry this might save you from scrounging
equipment or gas permeable covers.

Eric
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