Effect of repeated autoclavation
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jarlemag
Feb 5, 2013, 12:01:04 PM2/5/13
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Hello. A simple question popped up in the lab today, and I thought perhaps some of you would know: Will autoclaving the same medium multiple times degrade the quality of common complex growth media such as LB?
Best regards,
Jarle Pahr
Best regards,
Jarle Pahr
Daniel C.
Feb 5, 2013, 1:30:25 PM2/5/13
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On Tue, Feb 5, 2013 at 12:01 PM, jarlemag <jarle...@gmail.com> wrote:
> Hello. A simple question popped up in the lab today, and I thought perhaps
> some of you would know: Will autoclaving the same medium multiple times
> degrade the quality of common complex growth media such as LB?
I can't speak to the effects on LB, but repeatedly autoclaving growth
> Hello. A simple question popped up in the lab today, and I thought perhaps
> some of you would know: Will autoclaving the same medium multiple times
> degrade the quality of common complex growth media such as LB?
medium for Saccharomyces cerevisiae (yeast) can actually improve it by
causing precipitation of chemicals that would otherwise form
particulates and cloud up your final product. Because the important
chemicals are essentially sugar, autoclaving (even multiple times)
won't make the medium less suitable but it can make it progressively
darker in color.
-Dan
Avery louie
Feb 5, 2013, 1:54:20 PM2/5/13
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It can mess up the concentration by removing water, which makes it darker as has been mentioned.
--A
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Cathal Garvey
Feb 5, 2013, 2:02:25 PM2/5/13
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For LB, this isn't so much of an issue, but for media that contain
sugars you can get caramelisation of the sugar, splitting of
disaccharides, or maillard reactions, all of which might interfere with
some sensitive experiments, bacteria, or what have you.
For most experiments though, it won't make much of a difference provided
you try to use the same media batch for everything; doing otherwise, you
might have trouble telling what's different between samples; whether to
blame the bugs or the broth.
On 05/02/13 18:54, Avery louie wrote:
> It can mess up the concentration by removing water, which makes it
> darker as has been mentioned.
>
> --A
>
> On Tue, Feb 5, 2013 at 1:30 PM, Daniel C. <dcroo...@gmail.com
> <mailto:dcroo...@gmail.com>> wrote:
>
> On Tue, Feb 5, 2013 at 12:01 PM, jarlemag <jarle...@gmail.com
> <mailto:jarle...@gmail.com>> wrote:
> > Hello. A simple question popped up in the lab today, and I thought
> perhaps
> > some of you would know: Will autoclaving the same medium multiple
> times
> > degrade the quality of common complex growth media such as LB?
>
> I can't speak to the effects on LB, but repeatedly autoclaving growth
> medium for Saccharomyces cerevisiae (yeast) can actually improve it by
> causing precipitation of chemicals that would otherwise form
> particulates and cloud up your final product. Because the important
> chemicals are essentially sugar, autoclaving (even multiple times)
> won't make the medium less suitable but it can make it progressively
> darker in color.
>
> -Dan
>
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sugars you can get caramelisation of the sugar, splitting of
disaccharides, or maillard reactions, all of which might interfere with
some sensitive experiments, bacteria, or what have you.
For most experiments though, it won't make much of a difference provided
you try to use the same media batch for everything; doing otherwise, you
might have trouble telling what's different between samples; whether to
blame the bugs or the broth.
On 05/02/13 18:54, Avery louie wrote:
> It can mess up the concentration by removing water, which makes it
> darker as has been mentioned.
>
> --A
>
> On Tue, Feb 5, 2013 at 1:30 PM, Daniel C. <dcroo...@gmail.com
> <mailto:dcroo...@gmail.com>> wrote:
>
> On Tue, Feb 5, 2013 at 12:01 PM, jarlemag <jarle...@gmail.com
> <mailto:jarle...@gmail.com>> wrote:
> > Hello. A simple question popped up in the lab today, and I thought
> perhaps
> > some of you would know: Will autoclaving the same medium multiple
> times
> > degrade the quality of common complex growth media such as LB?
>
> I can't speak to the effects on LB, but repeatedly autoclaving growth
> medium for Saccharomyces cerevisiae (yeast) can actually improve it by
> causing precipitation of chemicals that would otherwise form
> particulates and cloud up your final product. Because the important
> chemicals are essentially sugar, autoclaving (even multiple times)
> won't make the medium less suitable but it can make it progressively
> darker in color.
>
> -Dan
>
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Sebastian S. Cocioba
Feb 5, 2013, 4:19:46 PM2/5/13
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It would cause a degradation to the agar for sure. Aside from that I've heard some speculation that the proteins in the LB may complex an become unavailable. I know for plant tissue culture MS does not fair well to multi-claves. Not sure exactly what happens but growth is stunted and the evidence points to nutrient deficiency so maybe the ions complex out of solution. Either way without a doubt agar will be messed up.
Sebastian S Cocioba
CEO & Founder
New York Botanics, LLC
Sent via Mobile E-Mail
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Nathan McCorkle
Feb 5, 2013, 7:31:41 PM2/5/13
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On Tue, Feb 5, 2013 at 10:54 AM, Avery louie <inact...@gmail.com> wrote:
> It can mess up the concentration by removing water, which makes it darker as
> has been mentioned.
The concentration could be fixed by just adding sterile distilled
> It can mess up the concentration by removing water, which makes it darker as
> has been mentioned.
water to some common fill line (maybe a mark that was draw before
inital heating)
I bet the darkening is more from glycation end products or the maillard reaction
--
-Nathan
Robert Sidney Cox III
Feb 6, 2013, 12:49:58 AM2/6/13
to diy...@googlegroups.com, cathal...@cathalgarvey.me
Glycerol will create Glyceraldehyde, which is toxic
> <mailto:diybio%2Bu...@googlegroups.com>. For more options,
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Cathal Garvey
Feb 6, 2013, 6:11:23 AM2/6/13
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Good to know, I had never heard that! Pretty relevant to luminescence
broth if anyone's doing bioluminescent bacterial isolations.
On 06/02/13 05:49, Robert Sidney Cox III wrote:
> Glycerol will create Glyceraldehyde, which is toxic
>
> On Wednesday, February 6, 2013 4:02:25 AM UTC+9, Cathal Garvey wrote:
>
> For LB, this isn't so much of an issue, but for media that contain
> sugars you can get caramelisation of the sugar, splitting of
> disaccharides, or maillard reactions, all of which might interfere with
> some sensitive experiments, bacteria, or what have you.
>
> For most experiments though, it won't make much of a difference
> provided
> you try to use the same media batch for everything; doing otherwise,
> you
> might have trouble telling what's different between samples; whether to
> blame the bugs or the broth.
>
> On 05/02/13 18:54, Avery louie wrote:
> > It can mess up the concentration by removing water, which makes it
> > darker as has been mentioned.
> >
> > --A
> >
> > On Tue, Feb 5, 2013 at 1:30 PM, Daniel C. <dcroo...@gmail.com
> <javascript:>
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broth if anyone's doing bioluminescent bacterial isolations.
On 06/02/13 05:49, Robert Sidney Cox III wrote:
> Glycerol will create Glyceraldehyde, which is toxic
>
> On Wednesday, February 6, 2013 4:02:25 AM UTC+9, Cathal Garvey wrote:
>
> For LB, this isn't so much of an issue, but for media that contain
> sugars you can get caramelisation of the sugar, splitting of
> disaccharides, or maillard reactions, all of which might interfere with
> some sensitive experiments, bacteria, or what have you.
>
> For most experiments though, it won't make much of a difference
> provided
> you try to use the same media batch for everything; doing otherwise,
> you
> might have trouble telling what's different between samples; whether to
> blame the bugs or the broth.
>
> On 05/02/13 18:54, Avery louie wrote:
> > It can mess up the concentration by removing water, which makes it
> > darker as has been mentioned.
> >
> > --A
> >
> > On Tue, Feb 5, 2013 at 1:30 PM, Daniel C. <dcroo...@gmail.com
> > <mailto:dcroo...@gmail.com <javascript:>>> wrote:
> >
> > On Tue, Feb 5, 2013 at 12:01 PM, jarlemag <jarle...@gmail.com
> <javascript:>
> >
> > On Tue, Feb 5, 2013 at 12:01 PM, jarlemag <jarle...@gmail.com
> > <mailto:jarle...@gmail.com <javascript:>>> wrote:
> > > Hello. A simple question popped up in the lab today, and I
> thought
> > perhaps
> > > some of you would know: Will autoclaving the same medium
> multiple
> > times
> > > degrade the quality of common complex growth media such as LB?
> >
> > I can't speak to the effects on LB, but repeatedly autoclaving
> growth
> > medium for Saccharomyces cerevisiae (yeast) can actually
> improve it by
> > causing precipitation of chemicals that would otherwise form
> > particulates and cloud up your final product. Because the
> important
> > chemicals are essentially sugar, autoclaving (even multiple
> times)
> > won't make the medium less suitable but it can make it
> progressively
> > darker in color.
> >
> > -Dan
> >
> > --
> > -- You received this message because you are subscribed to the
> > Google Groups DIYbio group. To post to this group, send email to
> > diy...@googlegroups.com <javascript:>
> > > Hello. A simple question popped up in the lab today, and I
> thought
> > perhaps
> > > some of you would know: Will autoclaving the same medium
> multiple
> > times
> > > degrade the quality of common complex growth media such as LB?
> >
> > I can't speak to the effects on LB, but repeatedly autoclaving
> growth
> > medium for Saccharomyces cerevisiae (yeast) can actually
> improve it by
> > causing precipitation of chemicals that would otherwise form
> > particulates and cloud up your final product. Because the
> important
> > chemicals are essentially sugar, autoclaving (even multiple
> times)
> > won't make the medium less suitable but it can make it
> progressively
> > darker in color.
> >
> > -Dan
> >
> > --
> > -- You received this message because you are subscribed to the
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> <mailto:diy...@googlegroups.com <javascript:>>. To
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> more options,
> > visit this group at
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> <https://groups.google.com/d/forum/diybio?hl=en>
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Matt Lawes
Feb 6, 2013, 8:07:38 AM2/6/13
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Yep the effect on sugars (heat hydrolysis) and Millard reaction (primarily with peptides and amino acids) of autoclaving - especially repeated autoclaving. I've always made concentrated sugar solutions, filter sterilized, and aseptically added to media after autoclaving.
>matt
Sent from my Verizon Wireless 4G LTE DROID
>matt
Sent from my Verizon Wireless 4G LTE DROID
-----Original message-----
Cathal Garvey
Feb 6, 2013, 8:12:12 AM2/6/13
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Had known that of sugars (although maillard is amino:sugar reaction, so
you can autoclave sugars separately and avoid this if a little bit of
caramelisation is OK), but not of glycerol.
Autoclaving: the quick and dirty sterilisation solution. :)
On 06/02/13 13:07, Matt Lawes wrote:
> Yep the effect on sugars (heat hydrolysis) and Millard reaction
> (primarily with peptides and amino acids) of autoclaving - especially
> repeated autoclaving. I've always made concentrated sugar solutions,
> filter sterilized, and aseptically added to media after autoclaving.
>
>>matt
>
> /Sent from my Verizon Wireless 4G LTE DROID/
>
>
> -----Original message-----
>
> *From: *Cathal Garvey <cathal...@cathalgarvey.me>*
> To: *"diy...@googlegroups.com" <diy...@googlegroups.com>*
> Sent: *Wed, Feb 6, 2013 06:11:31 EST*
> Subject: *Re: [DIYbio] Effect of repeated autoclavation
> > Learn more at www.diybio.org <http://www.diybio.org>
you can autoclave sugars separately and avoid this if a little bit of
caramelisation is OK), but not of glycerol.
Autoclaving: the quick and dirty sterilisation solution. :)
On 06/02/13 13:07, Matt Lawes wrote:
> Yep the effect on sugars (heat hydrolysis) and Millard reaction
> (primarily with peptides and amino acids) of autoclaving - especially
> repeated autoclaving. I've always made concentrated sugar solutions,
> filter sterilized, and aseptically added to media after autoclaving.
>
>>matt
>
>
>
> -----Original message-----
>
> *From: *Cathal Garvey <cathal...@cathalgarvey.me>*
> To: *"diy...@googlegroups.com" <diy...@googlegroups.com>*
> Sent: *Wed, Feb 6, 2013 06:11:31 EST*
> Subject: *Re: [DIYbio] Effect of repeated autoclavation
> > ---
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Matt Lawes
Feb 6, 2013, 8:21:53 AM2/6/13
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Yep, autoclaving sugar solution separately works if you don't overdo it (or forget about the bottle and leave it in there after the autoclave run is finished).
>matt
Sent from my Verizon Wireless 4G LTE DROID
-----Original message-----
From: Cathal Garvey <cathal...@cathalgarvey.me>
To: "diy...@googlegroups.com" <diy...@googlegroups.com>
&g
jarlemag
Feb 6, 2013, 10:14:41 AM2/6/13
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Very interesting replies. Thanks, everyone!
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