question
Shalini selvam
Hello there!To anyone concern:
I have a question.
How long can a media last after being autoclaved and where should we store it?can we autoclave it twice?
And how do we set up microbial consortia?
Thank.
Regards,Shalini
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John Gentile
Generally media that has been dispensed into screw capped tubes will
last a lot longer, especially if the caps are screwed tight after the
media has cooled. Petri dish media dries out and needs to be sealed in
a bag or container and usually requires refrigeration.
If you add blood to a media, it will shorten the "expriation date"
significantly. I wouldn't trust blood media past 4 to 6 weeks.
If you make the media in large flasks, you must cool the agar to about
50C and then pour into whaterver vial or plate you intend to use. It is
not recommended to autoclave again since that could carmalize some of
the sugars and may change some chemical properties and pH. You can
remelt media by bringing it above the melting temp of agar which is
around 97C, but don't go over that, just enough to get it all to melt.
There are a few good references for media preparation including the
Difco Manual and the American Society of Micribiology Manual of
Clinical Micribiology. These are the references I've used in my many
years of media making and storing.
--
John Gentile MS, M(ASCP)
Laboratory Information Mgr.
VA Medical Center
Providence, RI
yjg...@cox.net
Shalini selvam
Hello there!
Thank you Mr John and Bob for the previous reply.
Regarding the question on how to construct microbial consortia, im trying to
construct microbial consortia for bioremediation of hydrocarbon
contaminated soil. All the papers that i came across grew the inoculum
separately and then combined them.
Can we combine them directly and then test for the abiltiy of bacteria to degrade oil ? Eg lets
say flask 1 contains 1ml oil+100ml media+2ml inoculum of one type and
flask 2 contains 1ml oil+100ml media+ 1ml each of 2 different inoculum
for the testing?
Thank you!
Sincerely,Shalini
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Deirdre Sholto Douglas
>
> Hello there!
>
> Thank you Mr John and Bob for the previous reply.
>
> Regarding the question on how to construct microbial consortia, im trying to
> construct microbial consortia for bioremediation of hydrocarbon
> contaminated soil. All the papers that i came across grew the inoculum
> separately and then combined them.
What microbes comprise your consortium? Or rather, which are
you planning to use? Contrary to popular belief, microbes aren't
plug-and-play, things like competition for e- donors/acceptors
comes into play once you leave the realm of the Pure Culture...
there's considerably more to the process than simply bunging
bugs in a bottle and awaiting results.
> Can we combine them directly and then test for the abiltiy of bacteria to degrade oil ? Eg lets
> say flask 1 contains 1ml oil+100ml media+2ml inoculum of one type and
> flask 2 contains 1ml oil+100ml media+ 1ml each of 2 different inoculum
> for the testing?
Hydrocarbons (which includes the volatile organics) in general
or oil, in particular? And if only oil, then what sort of oil? Crude,
fuel, clean, dirty, high or low sulphur content?
IMO, the experiment you propose has a lot of variables which have
to be wrangled into submission before you'll get anything like a
reproducible result. I wish you the best, however.
Deirdre