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InvisibleImperfect Iam
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Agar solidified in wine bottle(unavoidable)
    #19334100 - 12/27/13 06:56 PM (10 years, 1 month ago)

I made some agar today and got everything ready, then I had to go to a family get together, I had forgotten all about. I made up the agar and then (after it had cooled some and pressure was nill, I had just opened PC and put in the bottle, I get a phone call telling me I was supposed to have been my aunts for Christmas 30 minutes ago.
So I sprayed some Lysol in the box and put cd sleeves into arm holes, now I am wondering which route to go as I read both searching through the archives.
Do I just put it in PC and boil it till it turns to a liquid, or do I re PC for ? Long. It's already been PC once fo 30 min. Do I re PC for ? Long, as I did not use any cotton or polyfill in original PC (I never do and have ran into 0 problems until now. I am fairly positive it is cont. free still, but do I risk the whole bottle by just boiling it having only A.F. on top or do I re PC for ?min.
All advice needed, I would say just boil it but w/o the polyfill stopper I am thinking I need to re PC idk, help me out here please And thank you!


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All you touch, and all you see, is all your life will ever be- Pink Floyd

Life is what happens while you're busy making other plans- John Lennon


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Offlinemonoculture
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Re: Agar solidified in wine bottle(unavoidable) [Re: Imperfect Iam]
    #19334111 - 12/27/13 07:00 PM (10 years, 1 month ago)

Would it hurt to PC again? I would just be sure and PC again, as you still have to heat it up again, it would be a little effort.


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OfflineCMOS
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Re: Agar solidified in wine bottle(unavoidable) [Re: Imperfect Iam]
    #19334122 - 12/27/13 07:02 PM (10 years, 1 month ago)

I usually just reheat it in the pc till its liquid again.  Actually doing that right now.


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InvisibleImperfect Iam
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Re: Agar solidified in wine bottle(unavoidable) [Re: monoculture]
    #19334144 - 12/27/13 07:11 PM (10 years, 1 month ago)

I don't think so as I saw a guy who did some for 90min. And it looked ok , but I thought I read stamets saying over cooking it could cause genetic mutations, if the malt caramelizes in mya.ha to the guy who just wrote in what a coincidence, did you use something other than A. F., if I would have used something else Along with theA.F., I wouldn't have a second thought on just boiling it!


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All you touch, and all you see, is all your life will ever be- Pink Floyd

Life is what happens while you're busy making other plans- John Lennon


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OfflineCMOS
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Re: Agar solidified in wine bottle(unavoidable) [Re: Imperfect Iam]
    #19334344 - 12/27/13 08:16 PM (10 years, 1 month ago)

I think its pretty hard to actually caramelize the malts, but I don't know I've only done a few agar runs.  I do at 15psi for 40min.  Extra light malt extract.

Almost everytime I either have too much agar to use, or something comes up, so I have to wait till later to use it.  I just heat it back up in the PC in its original container ( I use 1 1/2 pint jars with SFDs cause whiskey bottles are usually a little to tall for my SAB)  Once its liquid, swirl it up and pour like normal. 

Whats A.F.?


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OfflineRogerRabbitM
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Re: Agar solidified in wine bottle(unavoidable) [Re: CMOS]
    #19334357 - 12/27/13 08:19 PM (10 years, 1 month ago)

You won't caramelize the malts at 250F, but if you didn't have a filter on the agar bottle, I'd heat it up and dump it out.  Start over.
RR


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InvisibleImperfect Iam
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Re: Agar solidified in wine bottle(unavoidable) [Re: RogerRabbit]
    #19334456 - 12/27/13 08:44 PM (10 years, 1 month ago)

I just had an idea that should have been a no brainer, as it is still in SAB, I could remove foil, stuff with cotton or polyfil, then take it to PC and boil, all should be good! Agree?


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All you touch, and all you see, is all your life will ever be- Pink Floyd

Life is what happens while you're busy making other plans- John Lennon


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Re: Agar solidified in wine bottle(unavoidable) [Re: RogerRabbit]
    #19334466 - 12/27/13 08:45 PM (10 years, 1 month ago)

When I was a wee youngster doing the gruntwork in a big lab, I'd sometimes have to pour 100's of plates in a single day.  I would mix and autoclave all the flasks at once, then just dump them into a 40C water bath to keep them from solidifying.
A couple times, I forgot one on a benchtop and it would set up on me; a quick plop in the water bath and it was good to go again.
Never worried about filters, just foiled over the top of the flasks and made sure to give them a good spray with 70% EtOH before putting them in the hood to work.


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InvisibleImperfect Iam
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Re: Agar solidified in wine bottle(unavoidable) [Re: CliftonGK1]
    #19334516 - 12/27/13 08:56 PM (10 years, 1 month ago)

Good to know, but that was a lab setting, which my kitchen Is far from! Either way I will prob.be ok, but to be on the safe side...,


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All you touch, and all you see, is all your life will ever be- Pink Floyd

Life is what happens while you're busy making other plans- John Lennon


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Re: Agar solidified in wine bottle(unavoidable) [Re: Imperfect Iam]
    #19334536 - 12/27/13 09:01 PM (10 years, 1 month ago)

our lab keeps solidified agar in bottles, and microwaves them to remelt it and pour plates. We keep it like this for months at a time. Just have to be careful to microwave on low setting (10% power) so you don't explode it, since the bottom can melt and boil first... causing the solidified agar on top to violently exit the bottle.


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OfflineCliftonGK1
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Re: Agar solidified in wine bottle(unavoidable) [Re: kaptanoblivious]
    #19334557 - 12/27/13 09:08 PM (10 years, 1 month ago)

Quote:

kaptanoblivious said:
our lab keeps solidified agar in bottles, and microwaves them to remelt it and pour plates. We keep it like this for months at a time. Just have to be careful to microwave on low setting (10% power) so you don't explode it, since the bottom can melt and boil first... causing the solidified agar on top to violently exit the bottle.




I messed up once and did that with a bottle of ethidium bromide for pouring gel boxes.  We just thought it best to buy a new microwave for the lab instead of trying to clean that crap up.


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InvisibleImperfect Iam
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Re: Agar solidified in wine bottle(unavoidable) [Re: Imperfect Iam]
    #19334587 - 12/27/13 09:19 PM (10 years, 1 month ago)

Well I guess I am one lazy fuck as I am just boiling it w/o removing A.L. Foil to insert cotton, I will post in a few days and let everyone know if it all looks ok.
I hope I am not saying in a few days, you noob dumb fuck! Oh well what's a few plates and a little agar mix.
A lot if you are broke but I am fortunate enough to not be ATM!
And lol to Clifton, but ATM it prob. Wasn't so funny in your shoes!


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All you touch, and all you see, is all your life will ever be- Pink Floyd

Life is what happens while you're busy making other plans- John Lennon


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Re: Agar solidified in wine bottle(unavoidable) [Re: Imperfect Iam]
    #19334598 - 12/27/13 09:26 PM (10 years, 1 month ago)

I just rethought my stance on that,I am going to go ahead and re PC for 20minutes just to be safe also the I told you so I could feel coming my way!


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All you touch, and all you see, is all your life will ever be- Pink Floyd

Life is what happens while you're busy making other plans- John Lennon


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Re: Agar solidified in wine bottle(unavoidable) [Re: Imperfect Iam]
    #19334699 - 12/27/13 10:04 PM (10 years, 1 month ago)

So no nice pics of plates with all kinds of life on them soon? :tongue2:


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Re: Agar solidified in wine bottle(unavoidable) [Re: monoculture]
    #19334741 - 12/27/13 10:16 PM (10 years, 1 month ago)

Lol, EXACTLY


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All you touch, and all you see, is all your life will ever be- Pink Floyd

Life is what happens while you're busy making other plans- John Lennon


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Re: Agar solidified in wine bottle(unavoidable) [Re: Imperfect Iam]
    #19335743 - 12/28/13 06:23 AM (10 years, 1 month ago)

Let your plates sit for 7 days before using them if you're worried about potential contamination.  If they're contaminated, it will usually pop up inside of a week.


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Re: Agar solidified in wine bottle(unavoidable) [Re: CliftonGK1]
    #19338339 - 12/28/13 07:17 PM (10 years, 1 month ago)

Yeah, that's what I planned on thanks! I can't get over you worked in a lab, that would be one of my dream jobs, but I am sure they all have there drawbacks!


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All you touch, and all you see, is all your life will ever be- Pink Floyd

Life is what happens while you're busy making other plans- John Lennon


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Re: Agar solidified in wine bottle(unavoidable) [Re: CliftonGK1]
    #19338457 - 12/28/13 07:40 PM (10 years, 1 month ago)

Quote:

CliftonGK1 said:
Let your plates sit for 7 days before using them if you're worried about potential contamination.  If they're contaminated, it will usually pop up inside of a week.



This is great advice.  I have been quarantining every agar that I have done for at least 3-5 days before use.  It has only helped one time, but knowing that nothing but patience preserved a culture or two makes me feel good.


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Re: Agar solidified in wine bottle(unavoidable) [Re: Imperfect Iam]
    #19338472 - 12/28/13 07:43 PM (10 years, 1 month ago)

Quote:

cc1 said:
Yeah, that's what I planned on thanks! I can't get over you worked in a lab, that would be one of my dream jobs, but I am sure they all have there drawbacks!




Many labs over the course of 20+ years.  Everything from glass wash and garbage autoclaver, to class 10 cleanrooms, to BSL-3 containment labs, constant flow bioreactors, and even did a stint as a production engineer.
It's not as glam as it sounds.  Sure, I learned a hell of a lot; but there's a lot more days spent in a fluorescent microscopy darkroom analyzing 1000's of multiwells for evidence that an experiment worked (only to find out that it didn't) than there are days showing off at national conferences and feeling like you've actually accomplished something.
And production engineer is a fancy way of saying "nit-picky guy that all the production associates hate."


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Re: Agar solidified in wine bottle(unavoidable) [Re: HypnotoadCroaked]
    #19338523 - 12/28/13 07:52 PM (10 years, 1 month ago)

Quote:

Whippy said:
Quote:

CliftonGK1 said:
Let your plates sit for 7 days before using them if you're worried about potential contamination.  If they're contaminated, it will usually pop up inside of a week.



This is great advice.  I have been quarantining every agar that I have done for at least 3-5 days before use.  It has only helped one time, but knowing that nothing but patience preserved a culture or two makes me feel good.



Thank you! 

It's the same thing I used to do in the first clin micro lab I worked in.  When I started working in a production lab, all the sterile media we supplemented had to pass a 10 day hold before sale.  Prior to the hold, we did standard ANSI Z1.4 sampling on the production lot and cultured the sampled in TSB (tryp soy broth) at 37C to assure zero growth.

Working in a home lab with SAB poured plates, you're probably not going to have a 37C incubator and sterile TSB tubes to sample out a wedge and culture for a week and a half.  I sure as hell don't.  But, it's also not necessary when you're not dealing with the FDA and ISO audits.


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