[Microbiology] Re: Anaerobic culture technique

Lesley Robertson l.a.robertson at tnw.tudelft.nl
Mon May 15 03:35:44 EST 2006

"Matt McMahon" <matt.mcmahon at chee.queensu.ca> wrote in message 
news:mailman.975.1147646672.16885.microbio at net.bio.net...
>Hi all.  I'm looking for suggestions on anaerobic culture technique using 
>~100-150 mL serum bottles with butyl rubber stoppers.  I'm growing sulfate 
>reducing bacteria and previously had a nice system where I performed gas 
>exchanges on the bottle's headspace using a vacuum chamber hooked up to an 
>anaerobic gas cylinder.  This setup was nice because it was quick (handled 
>~20 bottles at a time) and allowed for good control by ensuring all bottles 
>in a batch received equal amounts of anaerobic gas.

>The vacuum chamber will no longer be available to me and I'm looking for 
>reasonably cheap alternative methods.  I know I could always manually gas 
>each bottle in a laminar flow hood.  This option is somewhat tedious and 
>lacks the previous degree of control.

I did a lot of work with denitrifiers and their enzymes using the anaerobic 
bags sold by Sigma, flushed with argon (much more effective than N2 or He. 
The only pain is the gloves fitted to the backs which seem to be desinged 
with the idea that only giants will use them - I tended to cut the gloves of 
and stick butyle gloves in their place. It was also easier to work after our 
workshop made me a work board to go into the bag to keep the work surface 
flat, otherwise it blew up like a balloon. Media, etc, went staight into the 
bag from the autoclave while still hot so that oxygen didn't have time to 
dissolve and thus didn't need removing.
This all worked fine, even with the most O2-sensitive denitrifiers.
Lesley Robertson
TU Delft, NL

More information about the Microbio mailing list

Send comments to us at biosci-help [At] net.bio.net