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Offlinekrypto2000
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Is pouring agar in a SAB safe? Doesn't the heat/steam cause all kins of airflow?
    #18864980 - 09/19/13 08:25 PM (10 years, 4 months ago)

I'm in the process of making some agar right now so just want to double check before pouring it. I know you have to pour it when it's still hot or else it will solidify. So why isn't this an issue? I'm not pouring it too soon am I? I basically pour it once it's cool enough to handle, should I be waiting even longer?

Also a side question. Yesterday I was told you're not actually supposed to use the plastic petri lids on the dishes as they allow contaminants in, which I've noticed to be the case in practice. I was told after nocing them up I can put them in plastic qt bags which are sterile, but what about storing them before I use them? Should they be safe if I pour them in the SAB and just let them sit in there until cool and then put them each in a separate plastic bag? That doesn't seem ideal, but I really can't think of a better idea personally. Will that work though?


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OfflineMush4Brains
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Re: Is pouring agar in a SAB safe? Doesn't the heat/steam cause all kins of airflow? [Re: krypto2000]
    #18865001 - 09/19/13 08:28 PM (10 years, 4 months ago)

You're definitely supposed to use the lids that come with the petris.  Open the dish, pour your agar, put the lid back on.  Let solidify and then put in a plastic bag.  The heat might cause a little bit of airflow but not enough to cause any damage.


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Re: Is pouring agar in a SAB safe? Doesn't the heat/steam cause all kins of airflow? [Re: Mush4Brains]
    #18865007 - 09/19/13 08:29 PM (10 years, 4 months ago)

I let mine cool in the SAB and then inoculate as soon as cool.


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Offlinekinkaku
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Re: Is pouring agar in a SAB safe? Doesn't the heat/steam cause all kins of airflow? [Re: tbagtag]
    #18865024 - 09/19/13 08:35 PM (10 years, 4 months ago)

I pour them then put them in bags in stackes of three or four and let them solidify and they're safe that way.


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Offlinekrypto2000
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Re: Is pouring agar in a SAB safe? Doesn't the heat/steam cause all kins of airflow? [Re: kinkaku]
    #18865204 - 09/19/13 09:12 PM (10 years, 4 months ago)

I was told you're supposed to remove the lid and use parafilm?


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Offlinekinkaku
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Re: Is pouring agar in a SAB safe? Doesn't the heat/steam cause all kins of airflow? [Re: krypto2000]
    #18865232 - 09/19/13 09:18 PM (10 years, 4 months ago)

Quote:

krypto2000 said:
I was told you're supposed to remove the lid and use parafilm?




by whom?


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Offlinekrypto2000
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Re: Is pouring agar in a SAB safe? Doesn't the heat/steam cause all kins of airflow? [Re: kinkaku]
    #18865265 - 09/19/13 09:24 PM (10 years, 4 months ago)

A quote from RR.

edit: actually upon re-reading the thread I think I misunderstood. You wrap the parafilm around the whole dish, lid included.


Edited by krypto2000 (09/19/13 09:25 PM)


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OfflineMush4Brains
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Re: Is pouring agar in a SAB safe? Doesn't the heat/steam cause all kins of airflow? [Re: krypto2000]
    #18865267 - 09/19/13 09:25 PM (10 years, 4 months ago)

I'm almost 100% positive he means wrap the entire dish, with the cover.


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Offlinekrypto2000
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Re: Is pouring agar in a SAB safe? Doesn't the heat/steam cause all kins of airflow? [Re: Mush4Brains]
    #18865269 - 09/19/13 09:26 PM (10 years, 4 months ago)

Yup, you're right, just edited my post to say that. :facepalm:


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OfflineMush4Brains
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Re: Is pouring agar in a SAB safe? Doesn't the heat/steam cause all kins of airflow? [Re: krypto2000]
    #18865277 - 09/19/13 09:27 PM (10 years, 4 months ago)

It's all good, at least you didn't try to leave the covers off before you poured and bagged. :lol:


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Offlinekrypto2000
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Re: Is pouring agar in a SAB safe? Doesn't the heat/steam cause all kins of airflow? [Re: Mush4Brains]
    #18865283 - 09/19/13 09:29 PM (10 years, 4 months ago)

So do you put the parafilm on inside the SAB too or are the dishes contained enough that it's safe to do so outside? I was going to use cling wrap as I don't have any parafilm atm. Not sure how the parafilm comes, but the clingwrap seems like it'd be hard to work with inside the SAB.


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Offlinekinkaku
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Re: Is pouring agar in a SAB safe? Doesn't the heat/steam cause all kins of airflow? [Re: krypto2000]
    #18865292 - 09/19/13 09:32 PM (10 years, 4 months ago)

the clingwrap or parafilm should be relatively sterile on the roll.


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OfflineMush4Brains
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Re: Is pouring agar in a SAB safe? Doesn't the heat/steam cause all kins of airflow? [Re: krypto2000]
    #18865480 - 09/19/13 10:23 PM (10 years, 4 months ago)

Quote:

krypto2000 said:
So do you put the parafilm on inside the SAB too or are the dishes contained enough that it's safe to do so outside? I was going to use cling wrap as I don't have any parafilm atm. Not sure how the parafilm comes, but the clingwrap seems like it'd be hard to work with inside the SAB.





You can cut the 1 foot rolls into 3 4" rolls with a sharp steak knife if you want it to be a little easier.


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Offlinekrypto2000
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Re: Is pouring agar in a SAB safe? Doesn't the heat/steam cause all kins of airflow? [Re: Mush4Brains]
    #18865539 - 09/19/13 10:39 PM (10 years, 4 months ago)

Well that was a pain in the fucking ass. The lid got steamed up and I couldn't tell if the dishes were full or empty. Ended up overfilling one and filling 2 with nothing because I couldn't tell the pyrex dish I pc'd the shit in was even empty. The overfilled dish ended up spilling all over my desk afterwards. Who knows if these things are contaminanted at this point or not, only ended up getting 3 dishes anyway and tossed 2 others because the shit got all over the outside. I'm not messing with agar again until I get some dishes that can actually go in the pressure cooker or a flow hood, fuck everything about this. I hate SAB work, causes more damn problems than it fixes. I feel like I might as well do mycology work handcuffed and blindfolded :thumbdown:


Edited by krypto2000 (09/19/13 10:41 PM)


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Offlinekinkaku
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Re: Is pouring agar in a SAB safe? Doesn't the heat/steam cause all kins of airflow? [Re: krypto2000]
    #18865554 - 09/19/13 10:43 PM (10 years, 4 months ago)

Quote:

krypto2000 said:
Well that was a pain in the fucking ass. The lid got steamed up and I couldn't tell if the dishes were full or empty. Ended up overfilling one and filling 2 with nothing because I couldn't tell the pyrex dish I pc'd the shit in was even empty. The overfilled dish ended up spilling all over my desk afterwards. Who knows if these things are contaminanted at this point or not, only ended up getting 3 dishes anyway and tossed 2 others because the shit got all over the outside. I'm not messing with agar again until I get some dishes that can actually go in the pressure cooker or a flow hood, fuck everything about this. I hate SAB work, causes more damn problems than it fixes. I feel like I might as well do mycology work handcuffed and blindfolded :thumbdown:




chillax,lol use an empty syring to pour the agar next time and also raise the lid enough so you can see whats going on.


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Offlinekrypto2000
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Re: Is pouring agar in a SAB safe? Doesn't the heat/steam cause all kins of airflow? [Re: kinkaku]
    #18865603 - 09/19/13 11:01 PM (10 years, 4 months ago)

So suck up the agar from my pyrex dish and then put it into the petris? Doesn't raising the lid kinda defeat the purpose of using a SAB? I guess the heat/steam would create an upward current though which would help prevent contaminant spores from falling into the work.


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Offlinekinkaku
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Re: Is pouring agar in a SAB safe? Doesn't the heat/steam cause all kins of airflow? [Re: krypto2000]
    #18865610 - 09/19/13 11:03 PM (10 years, 4 months ago)

I use a syringe to suck up agar then squirt it into the dishes. no mess and you dont have to raise the lid to high :shrug: but i do like to raise the lid high enough to see in.


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Offlinekrypto2000
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Re: Is pouring agar in a SAB safe? Doesn't the heat/steam cause all kins of airflow? [Re: kinkaku]
    #18865669 - 09/19/13 11:20 PM (10 years, 4 months ago)

That seems like a much better idea since then you can also measure exactly how much goes in each dish. Are the syringes hard to clean out afterwards since the agar is so sticky? Does the standard sucking up boiling water method still work just as well or are the syringes best not used for spore inoculation after that?


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Offlinekinkaku
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Re: Is pouring agar in a SAB safe? Doesn't the heat/steam cause all kins of airflow? [Re: krypto2000]
    #18865692 - 09/19/13 11:26 PM (10 years, 4 months ago)

actuaaly if you dont clean them before they solidify you just have to stick the whole tip of the needle in the boiling water to melt the agar then you just clean it like normal:thumbup:


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OfflineMush4Brains
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Re: Is pouring agar in a SAB safe? Doesn't the heat/steam cause all kins of airflow? [Re: kinkaku]
    #18865703 - 09/19/13 11:30 PM (10 years, 4 months ago)

If you get a large 140ml+ syringe + an extra long needle you can do many plates at once.


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Offlinekinkaku
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Re: Is pouring agar in a SAB safe? Doesn't the heat/steam cause all kins of airflow? [Re: Mush4Brains]
    #18865728 - 09/19/13 11:37 PM (10 years, 4 months ago)

Quote:

Mush4Brains said:
If you get a large 140ml+ syringe + an extra long needle you can do many plates at once.




never knew they made them that big :eek: (thats what she said)


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OfflineMush4Brains
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Re: Is pouring agar in a SAB safe? Doesn't the heat/steam cause all kins of airflow? [Re: kinkaku]
    #18865750 - 09/19/13 11:41 PM (10 years, 4 months ago)

Yeah, not only that, but then you can sterilize in a pint jar with SFD/Tyvek and a silicone port and just suck it all up after sterilizing without having to worry about not being sterile :trippinbawelz:


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Offlinekrypto2000
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Re: Is pouring agar in a SAB safe? Doesn't the heat/steam cause all kins of airflow? [Re: Mush4Brains]
    #18865789 - 09/19/13 11:49 PM (10 years, 4 months ago)

So you use the needle when 'pouring' the petris or do you take the needle off and just use the syringe? I thought it was the later, but now you guys are mentioning the needle.


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Offlinekinkaku
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Re: Is pouring agar in a SAB safe? Doesn't the heat/steam cause all kins of airflow? [Re: krypto2000]
    #18865796 - 09/19/13 11:53 PM (10 years, 4 months ago)

i do the needle because I do what mush4brains said and do my agar in a quart jar with filter patch and SHIP'S that way it stays nice and sterile.


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Offlinekrypto2000
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Re: Is pouring agar in a SAB safe? Doesn't the heat/steam cause all kins of airflow? [Re: kinkaku]
    #18865894 - 09/20/13 12:25 AM (10 years, 4 months ago)

Cool, well I'll try that next time then. I assume doing that I wouldn't really have much steam and thus wouldn't need to raise the lid either.


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Offlinekinkaku
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Re: Is pouring agar in a SAB safe? Doesn't the heat/steam cause all kins of airflow? [Re: krypto2000]
    #18865899 - 09/20/13 12:26 AM (10 years, 4 months ago)

:shrug: I've never had actual steam come off my agar other than the condensation on the lids after its poured.


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Offlinekrypto2000
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Re: Is pouring agar in a SAB safe? Doesn't the heat/steam cause all kins of airflow? [Re: kinkaku]
    #18865912 - 09/20/13 12:34 AM (10 years, 4 months ago)

Maybe I should have let it cool more? How long do you wait after the PCer is off and has released pressure? I took my dish out maybe 10-15 minutes after that and set it on the counter, waited maybe 20 minutes, so that the handle was cool enough to the touch, and then poured it.


Edited by krypto2000 (09/20/13 12:35 AM)


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Offlinekinkaku
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Re: Is pouring agar in a SAB safe? Doesn't the heat/steam cause all kins of airflow? [Re: krypto2000]
    #18865923 - 09/20/13 12:38 AM (10 years, 4 months ago)

I've never really timed it lol I just take the jar out of the pc when I can pick it up with a sock on my hand:shrug: then again I use the cheap one time use dishes so I dont have to sterilize those either.


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