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OfflinefirstTIMER420
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storing agar plates(fresh and innocd)
    #22055206 - 08/06/15 07:31 PM (8 years, 5 months ago)

Doing some agar tonight, and I really don't want to innoc them till the morning.  How do YOU store your fresh agar plates for a day or so? I was thinking about leaving them in the San and sealing the holes, having the plates wrapped with saran wrap or something.


For inoculated plates, I usually just put micropore tape on two sides so they close and stay closed and maybe put them in groups in ziplock bags.

But, I've been having problems with contamination. Not too terribly bad but a couple like 3-5 plates out of 20.(orange dots here and there).

Basically, what method do you use to store your plates?


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InvisiblebodhisattaMDiscordReddit
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Re: storing agar plates(fresh and innocd) [Re: firstTIMER420]
    #22055226 - 08/06/15 07:39 PM (8 years, 5 months ago)

put them un-wrapped but poured and cooled back into the sleeve they came out of. and twist the end shut.

or wrap the edges with saran wrap and put them on a shelf.


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InvisibleNumeroEno
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Re: storing agar plates(fresh and innocd) [Re: bodhisatta]
    #22055295 - 08/06/15 07:57 PM (8 years, 5 months ago)

My SAB stays set up all the time and I just leave my poured plates in the SAB unwrapped until ready to use. If they need to leave the SAB they get wrapped.


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Re: storing agar plates(fresh and innocd) [Re: NumeroEno]
    #22055342 - 08/06/15 08:06 PM (8 years, 5 months ago)

Overnight they will be fine sitting in your box...

Whenever I pour plates I put mine in fresh ziploc bags, 2 plates per.

Then use as needed when I'm ready for them.

After inoculating them I like to use parafilm but fresh bags or saran srap work too.


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OfflinefirstTIMER420
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Re: storing agar plates(fresh and innocd) [Re: PirateSwazey]
    #22056126 - 08/06/15 11:36 PM (8 years, 5 months ago)

And I'm assuming you guys wrap your plates after you noc them too?


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Re: storing agar plates(fresh and innocd) [Re: firstTIMER420]
    #22056497 - 08/07/15 02:35 AM (8 years, 5 months ago)

I was told to tape a stack of two or three together and place in a ziploc bag.


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Re: storing agar plates(fresh and innocd) [Re: uninc4life2010]
    #22056739 - 08/07/15 05:37 AM (8 years, 5 months ago)

When are you seeing contamination? If its before inoculation, itis most likely getting in during your pour, if its after inoculation, then it is either your innoculant (why we make transfers to new plates, away from the contams) or a flaw in your technique somewhere.

I wrap my plates with parafilm or a 1.5" cut off the end of a roll of cling wrap. Uninnoculted plates can be placed im short stacks of fresh ziplocks (theyre sterile) if they will be left somewhere they wont get knocked over.


IME bacteria in the plates after a pour is almost always from a sloppy pour, not getting in during storage


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Edited by mushpunx (08/07/15 05:41 AM)


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InvisibleNumeroEno
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Re: storing agar plates(fresh and innocd) [Re: mushpunx]
    #22058673 - 08/07/15 01:24 PM (8 years, 5 months ago)

Part of the reason I never get contams on my blank plates is because I pour when the agar is still pretty hot. I just make tall stacks of poured plates and I only get condensation on the top 2. Where I am it's necessary to wrap every plate or they dry out really fast.


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Re: storing agar plates(fresh and innocd) [Re: NumeroEno]
    #22058947 - 08/07/15 02:30 PM (8 years, 5 months ago)

Quote:

NumeroEno said:
Part of the reason I never get contams on my blank plates is because I pour when the agar is still pretty hot. I just make tall stacks of poured plates and I only get condensation on the top 2. Where I am it's necessary to wrap every plate or they dry out really fast.



you should pour when your agar is at about 47C to minimize condensation, also if you put your plates back into the sleeve they came in you can store your poured but not inoculated plates without them drying out. the sleeve is sterile on the inside still and the perfect shape and size considering the dishes came out of it.



make a few bottles up in one PC cycle save your extra bottles in the fridge. Re-melt them with a microwave on medium power, after they're melted put them in a water bath to adjust your agar to 47C


Edited by Trusted cuItivator (08/07/15 02:31 PM)


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InvisibleNumeroEno
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Re: storing agar plates(fresh and innocd) [Re: bodhisatta]
    #22058975 - 08/07/15 02:35 PM (8 years, 5 months ago)

I'll have to try that. I always destroy my sleeves when I open them though. I like the ziploc bag idea. When I eventually build a hood I'll have to start doing that or being more careful about how I open my sleeves. For now leaving my blank plates stacked in the SAB works well enough.


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:gd_icon:  Let it grow! Let it grow! Greatly yield!  :gd_icon:
What shall we say, shall we call it by a name
As well to count the angels dancing on a pin
Water bright as the sky from which it came
And the name is on the earth that takes it in

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OfflinefirstTIMER420
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Re: storing agar plates(fresh and innocd) [Re: bodhisatta]
    #22061306 - 08/08/15 01:08 AM (8 years, 5 months ago)

thanks for the responses guys.  I guess for now, ill just pour agar when I plan on inoculating it that day as to minimize the chances for contams.  I want to build/acquire a flowhood, but you all know they can be quite costly.

also, when I tried putting the plates back in the wrapper, a bunch of condensation formed in the bag and started dripping, weird.(I poured them really hot because I had to go to work, probably why)

ill just leave them in the sab till inoculated which is usually shortly after because I like to pour at about 118F-122F, and condensation is minimal at that point, and they also harden almost immediately.


As far as bacterial and other contams, im on my second transfer from a MS syringe originally, and I will be the first one to admit my technique needs some adjusting...to say the least.. You don't get those fluid motions your first day...anyways, my second transfer, im still getting orange bacteria on a couple plates and what looks like a different bacteria on the majority.  If this is being caused by my technique id like to figure it out and eliminate whatever im doing wrong.


EDIT:
After inoculating the plates, im wondering if have two or three in a bag, and one of those being contaminated would make the other ones get contamed, seemed to happen but not sure if im giving too much credit to the contam.

My recent ones I wrapped every other one in saran wrap and placed them in bags of two, just incase of the above


Edited by firstTIMER420 (08/08/15 07:04 PM)


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Offlinemushpunx
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Re: storing agar plates(fresh and innocd) [Re: firstTIMER420]
    #22066524 - 08/09/15 10:23 AM (8 years, 5 months ago)

"EDIT:
After inoculating the plates, im wondering if have two or three in a bag, and one of those being contaminated would make the other ones get contamed, seemed to happen but not sure if im giving too much credit to the contam.

My recent ones I wrapped every other one in saran wrap and placed them in bags of two, just incase of the above"


After inoculation, (or anything past a blank plate) almost all of us wrap the plates around the edge with parafilm or saran. Storing in a clean ziplock, is mostly done with blank plates only until they are poured.

This could be part of your problem.

If you have pictures it would be a big help. Take a tiny rice sized tranfer from the healthiest spot farthest away from any visible bacteria or mold colonies. If you see bacteria on the second plate it could be your culture grew over the bacteria and you are transfering it as well.. it can be sneaky.
Definitly a good reason to transfer as soon as you see healthy growth on an inoculation plate.

If you are seeing bacteria on any blank plates then its most likely your pour


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OfflinefirstTIMER420
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Re: storing agar plates(fresh and innocd) [Re: mushpunx]
    #22066957 - 08/09/15 12:18 PM (8 years, 5 months ago)

hm, so you don't put in a ziplock after innocing just wrap it? makes sense I guess. ill wrap my plates now they are a few days old and looking really good.


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Re: storing agar plates(fresh and innocd) [Re: firstTIMER420]
    #22067029 - 08/09/15 12:35 PM (8 years, 5 months ago)

Just wrap all mine in saran wrap. Takes all of 5-8secs per plate.


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OfflinefirstTIMER420
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Re: storing agar plates(fresh and innocd) [Re: maddchef]
    #22067270 - 08/09/15 01:39 PM (8 years, 5 months ago)

After wrapping you just leave em out? Or still put them in bags


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Re: storing agar plates(fresh and innocd) [Re: firstTIMER420]
    #22067478 - 08/09/15 02:28 PM (8 years, 5 months ago)

For the average hobbyist. I recommend hanging a shelf up in a draft less corner of a clean room to do your agar work on. It minimizes contams greatly and you don't need a flow hood. Once you make a transfer just leave them on the shelf upside down so no moisture runs onto your agar.




I maybe got one contam or two but since I leave them upside down after the transfer I haven't gotten any contams

Just a suggestion if your only starting out. The draft less corner feels as if your working in a huge Sab plus no restrictions on your arms. Hell I don't even wear gloves or wash my hands before. It will help you get your technique down I 100% re commend.



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Re: storing agar plates(fresh and innocd) [Re: ICEMANOO9]
    #22067690 - 08/09/15 03:24 PM (8 years, 5 months ago)

Personally after wrapping I just stack them anywhere I have room.

They stay clean. Be sure to wipe em down well before use


--------------------
In the land of the blind, the one eyed man is king.

All mushrooms are edible, but some only once.....                     

                                        Easier than cakes

I do science and shit.

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OfflinefirstTIMER420
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Re: storing agar plates(fresh and innocd) [Re: maddchef]
    #22067831 - 08/09/15 03:46 PM (8 years, 5 months ago)

I can't wait to get off work so I can go home and finish wrapping them. I'm nervous they are going to get cobtamed now. I do have them in individual bags right now though. Gonna wrap with saran wrap.
I figured wiping the outside with alcohol would be pointless since they are sterile or are you talking about when you are going to use them to transfer or innoc something else


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