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OfflinePeterPumpkinEater
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Registered: 08/25/19
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IS there a justifiable *need* or *use* for peroxide in the glovebox environment?
    #27048329 - 11/20/20 04:00 AM (3 years, 2 months ago)

Just curious.
And while I am at it I want to ask a related question, regarding contaminated specimens.

So I have some cubecelium and well, the petri dish is quite contammed with pink contams (qurius what it is).

Can I clean this specimen up by dunking it, like RR says, in a bleach solution, or should I maybe use peroxide? I'm really not sure on this, and cleaning up my contaminated culture is pretty much key for me at the moment.

My glove box is doing wonders, really a good one, but still a virgin, to be used soon when I get my little scale to weigh out the agar and malt extract... :smile:


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OfflineThe Dalcassian
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Re: IS there a justifiable *need* or *use* for peroxide in the glovebox environment? [Re: PeterPumpkinEater]
    #27049089 - 11/20/20 02:59 PM (3 years, 2 months ago)

There is no need to use peroxide at all. Soapy water will do fine for cleaning your SAB.

Isopropyl alcohol and water are also good for sanitising.

I'm sure someone will correct me if I'm wrong but, I'm pretty sure, if it's pink you should toss it. Pink mold can be bad for your health.


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OfflineYeetusdeetus
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Registered: 11/23/19
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Re: IS there a justifiable *need* or *use* for peroxide in the glovebox environment? [Re: The Dalcassian] * 2
    #27049099 - 11/20/20 03:06 PM (3 years, 2 months ago)

Yeah don’t fuck with that pink shit just start over from spores if you don’t have anything else that’s salvageable


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Invisiblemushhead
Potato Devourer
I'm a teapot


Registered: 08/22/14
Posts: 2,215
Loc: Dimension X-124
Re: IS there a justifiable *need* or *use* for peroxide in the glovebox environment? [Re: Yeetusdeetus]
    #27049117 - 11/20/20 03:18 PM (3 years, 2 months ago)

Hot soapy water, alcohol wipe down at the most, and proper sterile technique.
nothing more nothing less.


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OfflineTweeq
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Re: IS there a justifiable *need* or *use* for peroxide in the glovebox environment? [Re: Yeetusdeetus]
    #27049120 - 11/20/20 03:20 PM (3 years, 2 months ago)

The question is really if there is a justifyable use for a glovebox imo.

Lose the gloves and you have yourself a mighty SAB :smile: The true champion of low tech sterile work.


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Invisiblemushhead
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Registered: 08/22/14
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Re: IS there a justifiable *need* or *use* for peroxide in the glovebox environment? [Re: Tweeq]
    #27049157 - 11/20/20 03:37 PM (3 years, 2 months ago)

:whathesaid:
Why have permanent gloves attached when you can just idk.
Clean your hands and wear disposable gloves.


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OfflinePeterPumpkinEater
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Re: IS there a justifiable *need* or *use* for peroxide in the glovebox environment? [Re: mushhead]
    #27049787 - 11/21/20 01:43 AM (3 years, 2 months ago)

So the difference between a glove box and SAB is the gloves?

Also what about dipping cultures into bleach or peroxide, to decontaminate them, prior to transfer?

RR mentioned something about dipping in bleach, in relation to wild specimens he collected for propagation.


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Invisiblealaskappalachian
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Registered: 10/22/19
Posts: 1,674
Loc: The 49th Dimension
Re: IS there a justifiable *need* or *use* for peroxide in the glovebox environment? [Re: PeterPumpkinEater]
    #27049798 - 11/21/20 02:05 AM (3 years, 2 months ago)

Peroxide weakens the myc.  Don't dip your transfers in peroxide or bleach.  Not only is it totally unnecessary and not a substitute for proper technique,  it weakens the myc instead of helping.  What RR was referring to was a very dilute solution of bleach for multiple "washes" before plating wild transfers of real dirty birds like wild polypores and such.  Neither peroxide or bleach has any place whatsoever in cultivating actives.  Not for cubes or woodlovers fresh out of a woodchips pile or anything.  Observing proper sterile technique and becoming comfortable/familiar with working in a SAB or in front of a hood is the only way to get clean cultures.  Also gloves create pressure and thus currents when used (glovebox gloves/sleeves,  I mean,  of course...) whereas holes allow for relief when you move slowly.  Also why bigger holes (I use 6" diameter holes) are better than smaller (avoiding the "piston effect").


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Invisiblealaskappalachian
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Registered: 10/22/19
Posts: 1,674
Loc: The 49th Dimension
Re: IS there a justifiable *need* or *use* for peroxide in the glovebox environment? [Re: alaskappalachian]
    #27049802 - 11/21/20 02:14 AM (3 years, 2 months ago)

Also,  if you want an opinion on plates,  it really helps to post pics.  Pink could be a pernicious and/or hazardous mold (as mentioned) or pink bacterial colonies.  Either way I'd start over.  Did you use spore solution to inoculate your plates, or start with a print?


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"First we build the tools, then they build us."

THE 49th MYCOJOURNAL: Exotics, Auroras, and Entities


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OfflinePeterPumpkinEater
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Re: IS there a justifiable *need* or *use* for peroxide in the glovebox environment? [Re: alaskappalachian]
    #27049814 - 11/21/20 02:58 AM (3 years, 2 months ago)

I tossed some spores from a print using a "clean (haha)" butter knife, into a tiny jar with an inoculation port filled water and kept it on my bathroom shelf for a few months, saw the spores getting fuzzy around the edges, because for a while they didn't appear to do anything... clumps of spores, I mean.

I will still try and salvage a tiny bit of cubensis mycelium from the pink dread.

Here is a pic of my contams, low quality but you can see the pink stuff I am guessing is bacteria, because its so watery (the agar itself looks almost too dry but I don't know)... I'm amazed at how slow the white mycelium is growing, I would have expected an overgrown plate after what, 10-14 days?, but you can see some "pure" bits of white mycelium growing that doesn't have any gunk in it: My question is if I could at least try and use an inoculation loop, steal off a tiny bit of that white mycelium, and maybe dunk it in diluted bleach to try and clean it a bit, to see at least if I can get a clean culture from this plate? :



Interesting about the gloves and pressure. Maybe I can create another hole with polyfill to act as some type of "depressurization filter" :p
Donno, but our house is literally encrusted with molds, the table my glovebox is on is ... almost blackened here and there due to mold that just grew into the filth, its on an old and neglected semi-outdoor patio that we don't really ever use...


btw your dancing leprechaun avatar, is it from "The Last Leprechaun"? Very fun movie on Tubi.tv:

https://tubitv.com/movies/528963/the-last-leprechaun


Edited by PeterPumpkinEater (11/21/20 03:01 AM)


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OfflineYeetusdeetus
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Registered: 11/23/19
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Re: IS there a justifiable *need* or *use* for peroxide in the glovebox environment? [Re: PeterPumpkinEater]
    #27049852 - 11/21/20 04:09 AM (3 years, 2 months ago)

The pic is kinda fuzzy but personally I wouldn’t touch that. You mentioned a “clean” knife, did you flame sterilize it until red hot or just wash it? Flaming your tool until red hot before each transfer is the best way to ensure nothing from the environment or previous transfers follows you to other plates.

I’d try to set up a SAB inside to get away from as many potential drafts as possible. Don’t do your sterile work on a moldy table, buy a cheap folding table you can use for sterile work if nothing else is available. You’ll never be able to stuff polyfill tight enough in a glovebox to make it impenetrable to contaminants

I know a lot of people tout being able to do their agar work in a moldy basement but people often omit how much better your technique will have to be in a dirty environment to achieve the same degree of success

I’d highly recommend buying one or two #7 scalpel handles and a pack of #11 blades. The blades are thinner which makes sterilizing go by much quicker.


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OfflinePeterPumpkinEater
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Re: IS there a justifiable *need* or *use* for peroxide in the glovebox environment? [Re: Yeetusdeetus]
    #27049919 - 11/21/20 05:58 AM (3 years, 2 months ago)

It was most certainly clean, but not sterile, I wiped it with a rag... I was at that stage under the impression that I wanted to try and cultivate mushrooms in an asterile fashion, believing that like nature, that it suredly must be possible: I have since been playing another fiddle and am doing it by the book from now on, definitely. I still have another clean spore print, that spore-water is definitely laced with vileness incarnate...!


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OfflineTweeq
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Re: IS there a justifiable *need* or *use* for peroxide in the glovebox environment? [Re: PeterPumpkinEater]
    #27050009 - 11/21/20 08:02 AM (3 years, 2 months ago)

https://www.shroomery.org/forums/showflat.php/Number/23990913

Here is Bod's SAB tek for you. He's got links to all the other teks you need to learn to grow in his signature too.

As you have probably already found out, copying nature doesn't work too well for cubes. In nature you don't see full canopies of them anywhere. For that you need sterile tek


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OfflinePeterPumpkinEater
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Re: IS there a justifiable *need* or *use* for peroxide in the glovebox environment? [Re: Tweeq]
    #27050480 - 11/21/20 01:38 PM (3 years, 2 months ago)

Thanks Tweeq! Shall take a gander at
his SAB in a sec.


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OfflineTweeq
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Re: IS there a justifiable *need* or *use* for peroxide in the glovebox environment? [Re: PeterPumpkinEater]
    #27050481 - 11/21/20 01:39 PM (3 years, 2 months ago)

Quote:

PeterPumpkinEater said:
Thanks Tweeq! Shall take a gander at
his SAB in a sec.




You're welcome!


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OfflinePeterPumpkinEater
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Re: IS there a justifiable *need* or *use* for peroxide in the glovebox environment? [Re: Tweeq]
    #27051487 - 11/22/20 04:12 AM (3 years, 2 months ago)

I designed my GB in the same way. Metal can idea to burn holes. I think tbh I would want to buy a tent and put a plastic San and sidetable in it as I could have maybe more controlled air in it? Our house is terrifyingly dusty and moldy, but I will be trying to use my GB as an SAB . Can just remove the gloves. I will work as quickly as possible when putting anything into what I know to be sterile agar Petros to reduce risk of exposure and reduce contam vectors


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OfflineTweeq
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Re: IS there a justifiable *need* or *use* for peroxide in the glovebox environment? [Re: PeterPumpkinEater]
    #27051495 - 11/22/20 04:54 AM (3 years, 2 months ago)

What can also help is to practice your movements before working on actual cultures. Just find the ideal places for everything you need so you can go with the least amount of movement and time since all your stuff is already placed where it should be.

Excuse my English. Hope it makes sense for you


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Offlineadhoc
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Re: IS there a justifiable *need* or *use* for peroxide in the glovebox environment? [Re: PeterPumpkinEater]
    #27051530 - 11/22/20 05:56 AM (3 years, 2 months ago)

Quote:

PeterPumpkinEater said:
I will work as quickly as possible when putting anything into what I know to be sterile agar Petros to reduce risk of exposure and reduce contam vectors




As Tweeq alluded to it's important to know what you plan to do every time you stick your hands into the SAB, and then do it at a relaxed pace with smooth movements.  Going quickly is more likely to stir up air currents.


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OfflinePeterPumpkinEater
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Re: IS there a justifiable *need* or *use* for peroxide in the glovebox environment? [Re: adhoc]
    #27058371 - 11/26/20 09:30 AM (3 years, 2 months ago)

definitely. a question. So if I keep the gloves mountaed on te side, maybe I can cut another ring-hole, and fill it with polyfill, to reduce the pressure?

I'd really prefer not to remove the gloves as I fear air might be sucked in. But maybe I should just be very careful...

Not sure what you guys think about the idea of adding another hole (to reduce the pressure of the mounted gloves), but cover it with polyfill, to make up for that pressure difference.


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OfflineTweeq
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Re: IS there a justifiable *need* or *use* for peroxide in the glovebox environment? [Re: PeterPumpkinEater]
    #27058516 - 11/26/20 10:53 AM (3 years, 2 months ago)

Idk. SAB is proven tech and simple and cheap af


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OfflineYeetusdeetus
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Re: IS there a justifiable *need* or *use* for peroxide in the glovebox environment? [Re: Tweeq]
    #27058519 - 11/26/20 10:54 AM (3 years, 2 months ago)

:whathesaid:



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