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OfflineMr.Meeseeks
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EPIC fail first try pouring plates * 1
    #26652386 - 05/06/20 09:22 AM (9 months, 20 days ago)

Hi all,

After making my agar mix last night I thought i would tackle pouring in my SAB today. A number of mistakes I made I thought i'd put up here for people to NOT repeat.

1. Waited till too late at night to start, and rushed the cooling/taping of the plates. I didn't let them cool in the stack because I thought i had to deal with the condensation straight away, which I did by the way OUTSIDE of my SAB.....
2. Sprayed so much soapy water on the front of my SAB inside so that I couldn't see what I was doing, wasted 4 plates because i didn't realise I had picked the lid off and then contaminated the insides
3. Poured way too much agar into each plate...

So anyway, I have 6 plates that might be ok, but the condensation keeps on coming back in all of them so I've obviously messed up the temperature. Can see lots of water beads on the inside of my agar and have read up on how this will affect my plate if I do try to streak it.

Anyways, I have more materials so I am going to try again tomorrow, not repeat my mistakes and take my time with it. Hope ya'll dont make the same mistakes I did.


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Re: EPIC fail first try pouring plates [Re: Mr.Meeseeks]
    #26652442 - 05/06/20 10:00 AM (9 months, 20 days ago)

You’ll always get some level of condensation, usually only on the top of the stack. When you pour them just leave them stacked in the SAB overnight or for a couple days and it’ll go away.

An easy way to learn when you start out is to take five plates in a stack, lift the lid of the bottom one up (with the 4 empty plates still on top of it) just enough to pour the agar in, then set the lid and stack back down.

Then do it again with the lid of the next empty plate and so on.

It’s very quick and easy, doesn’t expose much of the open plate, and keeps the fresh plates stacked so you can leave them to cool without trying to move them without spilling.

Sometimes I’ll take two old plates and fill them with hot water and add them to the top of the stack so they absorb most of the condensation and all the agar plates come out clear.


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OfflineTrippyMomp
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Re: EPIC fail first try pouring plates [Re: Mr.Meeseeks]
    #26652443 - 05/06/20 10:00 AM (9 months, 20 days ago)

Good luck. I'm messing with agar too rn. Had allot of the same issues as you. I'm gonna be trying to no pour next time. It may take a while in ny small pc but atleast I'll know I wont be contaminating or pouring to heavy or to quick with it. On a side note I havent tried yet but apprently a hot cup of liquid with a flat bottom can be briefly rested on top and will make the condensation resttle for a bit for viewing. Also just so you dont make the same mistake as me and ruin your plates well allot of them is make or buy a inoculation loop so you dont flood your plates. Allot of mine are flooded from the syringes and I think they had a decent amount of bacteria in them so Its making it harder to work with. The less you use to inoculate with using a dirty syringe the better. Maybe you have clean shit tho. Just didnt want you doing the same shit as me set me back a week or two.


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OfflineMr.Meeseeks
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Re: EPIC fail first try pouring plates [Re: TrippyMomp]
    #26653264 - 05/06/20 06:33 PM (9 months, 19 days ago)

good call on the stacking and leaving in SAB. I have a loop that I'm going to streak across the plate, dont have anything to flame it yet though, need to get one today. That being said my spore print process from wild fruits wasn't exactly a sterile process and some of them were done a year ago. Will have to just see how it goes, hopefully I'll be able to isolate something of use


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OfflineTrippyMomp
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Re: EPIC fail first try pouring plates [Re: Mr.Meeseeks]
    #26654741 - 05/07/20 11:39 AM (9 months, 19 days ago)

Quote:

Mr.Meeseeks said:
good call on the stacking and leaving in SAB. I have a loop that I'm going to streak across the plate, dont have anything to flame it yet though, need to get one today. That being said my spore print process from wild fruits wasn't exactly a sterile process and some of them were done a year ago. Will have to just see how it goes, hopefully I'll be able to isolate something of use



I use like a 10 dollar cigar torch. crown Brule torches if you wanna get boujee


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Re: EPIC fail first try pouring plates [Re: TrippyMomp]
    #26654759 - 05/07/20 11:47 AM (9 months, 19 days ago)

Yeah I got a culinary torch for like ten bucks it’s great.


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OfflineMr.Meeseeks
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Re: EPIC fail first try pouring plates [Re: A.k.a] * 2
    #26656241 - 05/08/20 12:30 AM (9 months, 18 days ago)

Lol I have a full on plumbing gas torch, woops


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OfflineLogicaL ChaosM
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Re: EPIC fail first try pouring plates [Re: Mr.Meeseeks]
    #26656438 - 05/08/20 03:11 AM (9 months, 18 days ago)

Great for sterilizing AND dabbing :awehigh:


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Offlinemushpunx
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Re: EPIC fail first try pouring plates [Re: LogicaL Chaos]
    #26656484 - 05/08/20 03:54 AM (9 months, 18 days ago)

Taping? What are you using to wrap your plates?

Also inoculated plates you can put in fresh ziplocks, they are sterile. I put a few in each one. When you pour at the right temperature you shouldn't get any condensation, even on the top plates. Bodhisattva has a great Tek about this.

Since you poured these outside the SAB, Chuck them out. Or watch them to see what happens first for fun but I wouldn't use them.


  Good luck on your next attempt!


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Edited by mushpunx (05/08/20 05:42 AM)


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OfflineMr.Meeseeks
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Re: EPIC fail first try pouring plates [Re: mushpunx]
    #26659241 - 05/09/20 06:56 AM (9 months, 17 days ago)

i poured them in the SAB but i took them out of the SAB to try and deal with the condensation and to tape them. just used a thin microfilm tape


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OfflineTrippyMomp
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Re: EPIC fail first try pouring plates [Re: Mr.Meeseeks]
    #26659459 - 05/09/20 10:14 AM (9 months, 17 days ago)

Quote:

Mr.Meeseeks said:
i poured them in the SAB but i took them out of the SAB to try and deal with the condensation and to tape them. just used a thin microfilm tape




That's ok just make sure you're storing them upside down. Put the unused in the fridge for later. Just wype them with iso before you put them back in the Sab to do work.


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Offlinemushpunx
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Re: EPIC fail first try pouring plates [Re: Mr.Meeseeks]
    #26660053 - 05/09/20 03:31 PM (9 months, 17 days ago)

Quote:

Mr.Meeseeks said:
i poured them in the SAB but i took them out of the SAB to try and deal with the condensation and to tape them. just used a thin microfilm tape




Ah I see.  Cut a 1" section of the end of a roll of plastic cling wrap with a sharp knife and use that to wrap the edges of your plates.

Or Parafilm if you can get it, that's what it's made for


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Offlinemushpunx
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Re: EPIC fail first try pouring plates [Re: TrippyMomp]
    #26660063 - 05/09/20 03:39 PM (9 months, 17 days ago)

Quote:

TrippyMomp said:
Quote:

Mr.Meeseeks said:
i poured them in the SAB but i took them out of the SAB to try and deal with the condensation and to tape them. just used a thin microfilm tape




That's ok just make sure you're storing them upside down. Put the unused in the fridge for later. Just wype them with iso before you put them back in the Sab to do work.





Why upside down?

I prefer to store uninoculated plates at room temp. That way you will see if they are sterile,you can see if anything grows on them


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OfflineTrippyMomp
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Re: EPIC fail first try pouring plates [Re: mushpunx]
    #26660169 - 05/09/20 04:35 PM (9 months, 16 days ago)

Quote:

mushpunx said:
Quote:

TrippyMomp said:
Quote:

Mr.Meeseeks said:
i poured them in the SAB but i took them out of the SAB to try and deal with the condensation and to tape them. just used a thin microfilm tape




That's ok just make sure you're storing them upside down. Put the unused in the fridge for later. Just wype them with iso before you put them back in the Sab to do work.





Why upside down?

I prefer to store uninoculated plates at room temp. That way you will see if they are sterile,you can see if anything grows on them




Might be right in that sense. I'm not sure at all I'm a noob. But I have seen all over the internet, and it's just because bacteria/ dust particles settle on the ground so if its upside down the thought is that it's a buffer and contams are less likely to get into the stuff but idk. Like I said I deff couod be wrong and would like to know standard procedure.also I live in a desert so in worried about the plates evaporating to quick out of the fridge but I'm not experienced at all.


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Re: EPIC fail first try pouring plates [Re: TrippyMomp]
    #26660175 - 05/09/20 04:37 PM (9 months, 16 days ago)

I'm not doing this with my glad twist locks tho. And its probably less damage keeping them right side up now that I think about it considering having them upside down makes me have to flip them to peak.


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Re: EPIC fail first try pouring plates [Re: TrippyMomp]
    #26660365 - 05/09/20 06:18 PM (9 months, 16 days ago)

I live in a desert also and my plates last a long time. I’ve only had a few dry out and they were soft and thin. If you’re pouring regular thickness/hardness and keeping them wrapped I could see them going a year no problem.

I’ve got a bunch of 5 month old plates thatve been on a desk for two months then in a drawer since then that are still in good condition.


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OfflineMr.Meeseeks
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Re: EPIC fail first try pouring plates [Re: A.k.a]
    #26660544 - 05/09/20 07:58 PM (9 months, 16 days ago)

good call. i will take them out of the fridge and see if any contams show up. storing upside down is an interesting idea, i feel like i read that somewhere on the forum too. I tried the water bath but i did it so late at night i rushed it and poured too hot, some of my plates dont have alot of condensation though they actually look ok. Still haven't heard back from the supplier if their MEA mix is MEA mixed with agar agar, you'd have to assume so though. cling film is a good call, probably easier to peel off than the tape i used. Looking forward to streaking the plates


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Re: EPIC fail first try pouring plates [Re: Mr.Meeseeks]
    #26660650 - 05/09/20 08:55 PM (9 months, 16 days ago)

Cling wrap is the best, I couldn’t imagine using tape.

Definitely cut the roll into smaller pieces though. I put off doing that forever and when I finally did cut it things were a million times easier.

It’s a lot faster and easier than you’d think.

Plus I’ve probably wrapped 1500 plates and only used two inches of the roll.


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