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OfflineEnvypenis
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70% iso doesn’t kill all bacteria
    #28023689 - 10/29/22 09:32 PM (2 years, 2 months ago)

Been experimenting with mycology for a little over a year now. It’s been a learning experience to say the least. This stuff makes growing world class weed look easy lol battled a lot already but my recent problem has to do with getting clean agar dishes. Had great success until a couple months ago.

I’ve improved on my pressure cooking technique. I do quart jars with new lids every time. Filled to within 1/2” headspace. I let steam blow out 10 mins to push out air of PC, then do a forty minute cook at 15 psi, wait for pressure to come down to 0 to remove. I get zero boil over, the lid is popped down and sealed before I even take the jar out of the PC. I have to assume my agar is safe as jams are done this way and will last for years on shelves.

I either wait for the agar to hit 130* or water bath until I’m ready. Have to bring it down to my tent, re sterilize everything and every time my hands leave that box for a second they get douched. Soak my lab Coat arms. I mean I can barely breathe in this tent with the alcohol fumes. Last batch i worked the stack of dishes from the bottom up. Still got half contaminated on the Amazon sterilized petri dishes. Had extra agar tried pouring into the jello Shots and got 60% clean - better than Petri dishes. Go figure.. 

I do all my sterile work inside a grow tent which I clean weekly with a bleach water mix. Stainless work table with air box from Amazon which has zippered clear vinyl sides, fabric bottom.. hepa 2500 sq ft filter inside the 4x8 tent. Positive pressurized - clean vented in with filtration 13 cloth material secured over air inlet. I’m always showered and no shoes in my tent. Clean clothes, lab coat with rubber bands over the cuffs with xtra long gloves up my wrists. I spray 70% iso all over the SAB. Soaking it to the point it’s dripping and the bottom is a puddle. I wipe and sanitize everything multiple times. I’ve tried brand new bagged and sealed x 2 manufactures disposable Jell-O shot cups as well as now the disposable but supposedly sterile PET petri dishes off Amazon. Still have contam all the way thru. Two batches ago 3 out of 25 jellos made it after a week sitting in my SAB.

Consistently the last three or four times I’ve poured I’m getting milky or cloudy spots within 24 hours on my agar. Some small raised shiny bacterial Spots as well. From what I’m reading in all the teks.. I’m way over killing it on sterilization.

Here is what I believe the issue is but I’d love to get some feedback and if anyone has experienced this…

70% iso is not killing all the bacteria in my SAB. According to these findings.. of 282 samples that were “disinfected” with 70% iso - 100 samples still contained microorganisms. Is it possible that I have an alcohol resistant bacteria in my SAB? Or am I losing my mind over something stupid that I’ve missed along the way here..

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4623738/

Edited by Envypenis (10/29/22 09:39 PM)

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OfflineBrewtality
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Re: 70% iso doesn’t kill all bacteria [Re: Envypenis] * 1
    #28023705 - 10/29/22 09:42 PM (2 years, 2 months ago)

Have you considered that it isn't a STERILE air box and instead a still air box. Are you turning the fan off when you go into the tent? Try slowing way down and trying not to disturb the air when working in the box. I have to remind my repeatedly to slow down or I my speed slowly increases. Have you tried making a video of your process. That might help you understand where things are going wrong.

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OfflineBajazly
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Re: 70% iso doesn’t kill all bacteria [Re: Brewtality]
    #28023711 - 10/29/22 09:46 PM (2 years, 2 months ago)

Positive pressurized? You have a fan running thru your SAB? Filtered?

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OfflineTheUsualSuspect
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Re: 70% iso doesn’t kill all bacteria [Re: Envypenis]
    #28023721 - 10/29/22 09:51 PM (2 years, 2 months ago)

Quote:

Envypenis said:
Had great success until a couple months ago.






What changed from when you were getting success to now? Must have been a process or a piece of equipment.

Im not sure I completely undestand your whole process but are you using the grow tent as a still air box but also pumping air into it?


Quote:

Envypenis said:
clean vented in with filtration 13 cloth material secured over air inlet"





that part isnt tracking well either.

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Offlinehazyhorse
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Re: 70% iso doesn’t kill all bacteria [Re: Envypenis] * 1
    #28023753 - 10/29/22 10:08 PM (2 years, 2 months ago)

you’re going way overboard dude. you can generally work in a dry, no prior cleaning SAB with bare hands & get perfect results. doesn’t hurt to be clean & wear gloves, but main thing is to work in a draft free room with no fans running. as someone else said, it’s a STILL air box, not STERILE air box. gravity is what keeps your media sterile, not clean air. you’d have to work in a lab grade HEPA filtered clean room wearing full PPE & shit for it to be sterile. you should not be gassing yourself out with chemical fumes man. you’re always going to have bacteria in your SAB because it’s not a sterile environment. as mentioned earlier, the air in the room being still & your motions being smooth, slow, & deliberate are going to make way more of a difference than bleaching out a grow tent & then running a fan.


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OfflineSingularFusion
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Re: 70% iso doesn’t kill all bacteria [Re: TheUsualSuspect] * 2
    #28023754 - 10/29/22 10:08 PM (2 years, 2 months ago)

so you say your SAB has clear plastic and zippers?

Well I bet the walls move a little, this causes a suction and blowing type effect, sucking contam in through wherever there is negative pressure, which will be the arm holes in this case I guess. Why don't you build a simple SAB from a stiff plastic tub, I would bet your situation would improve

SAB works on still air... The concept of sterilizing it with iso is asinine, iso only gives things a good clean, sterile is not expected. Using the SAB successfully is all about having good technique and a still room around you. If you are blowing air in to your workspace(filtered or not), you don't have a still air box anymore and as such it works like you have seen it work, poorly

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Offlinehazyhorse
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Re: 70% iso doesn’t kill all bacteria [Re: SingularFusion]
    #28023756 - 10/29/22 10:11 PM (2 years, 2 months ago)

:whathesaid:

i didn’t even realize it wasn’t a hard plastic SAB when i read that the first time. the flexible sheets are gonna stir up a ton of air even with subtle movements. that isn’t helping anything


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Offlinethe man
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Re: 70% iso doesn’t kill all bacteria [Re: TheUsualSuspect]
    #28023760 - 10/29/22 10:13 PM (2 years, 2 months ago)

your issue is not the 70% rather the air movement. turn off the fan while working. is there soil/hydro in this tent aswell? imagine thats part of ur issue to as dont want cloth/carpet nevermind soil in the room.

the KEY is still air NOT cleaning the air to 100% sterile. u can rub ur hands with strik 9 and bleach and if wave them over agar they will contam 60% of the time.

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OfflineTheUsualSuspect
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Re: 70% iso doesn’t kill all bacteria [Re: SingularFusion]
    #28023763 - 10/29/22 10:15 PM (2 years, 2 months ago)

All that slopping cleaning chemicals everywhere and gassing yourself out, id stop doing that.

You seem willing to spend a little money on your hobby. If I were you I would get a small flow hood unit and put it inside that grow tent (and close off the outside air flow vents) and then you have a solid foundation to do work.

Get in the tent, zip it closed, let the flow hood run a bit and then start the lab work.

just my opinion

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Offlinethe man
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Re: 70% iso doesn’t kill all bacteria [Re: TheUsualSuspect]
    #28023770 - 10/29/22 10:27 PM (2 years, 2 months ago)

naa def dont need to close off vents with flow hood. u want air to move out and away easily not making eddies and swirling things up in a tiny room. anyhoo. SAB still air box NO moving air near ur box or made by fast movements. simple.

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OfflineSirPsycho
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Re: 70% iso doesn’t kill all bacteria [Re: the man]
    #28023794 - 10/29/22 10:56 PM (2 years, 2 months ago)

No shit it doesn't kill all bacteria. It's a sanitizer, it doesn't sterilize.

An SAB is a STILL AIR box. Not a clean air box.


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InvisibleBaba Yaga
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Re: 70% iso doesn’t kill all bacteria [Re: the man] * 3
    #28023803 - 10/29/22 11:09 PM (2 years, 2 months ago)

You have this POS right?




This is the reason for your failure. You will be better off throwing this disgrace of SAB away. I can't believe those ass wipes are actually selling something like this and their advertising is making me throw up, "designed with you in mind", yeah right.

Sorry for the rant it's not your fault, but honestly get rid of this thing and build a SAB out of a rigid plastic tote. Reduce your alcohol use to a minimum and rely on good sterile technique......you know, the kind of technique you weren't able to achieve with this floppy SAB these dickheads sold to you.

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Offlinematsc
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Re: 70% iso doesn’t kill all bacteria [Re: Baba Yaga]
    #28023845 - 10/29/22 11:34 PM (2 years, 2 months ago)

While I do agree with what folks are saying about the Positive-pressuretent and SAB, and that you are going a bit overboard at times....

My microbiologist heart requires me to say that, yes, there are in fact some bacteria that can survive 70% isopropanol exposure. Hell, when we buy Bacillus endospores to play with in my lab, they're literally sent as an alcohol suspension!

But they are a fairly niche group, and exposure time matters, and etc etc etc. In general, rubbing alcohol is fine as a surface sanitizer. Won't totally sterilize stuff, but useful tool in most situations.

And all that said, when doing lazy lab stuff at home, I just make pre-pour plates. Yeah the condensation in them is a little annoying, but when I only need a few plates at a time it's just easier for me. And I am VERY lazy most of the time :grin:


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Offlinehazyhorse
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Re: 70% iso doesn’t kill all bacteria [Re: matsc]
    #28023850 - 10/29/22 11:38 PM (2 years, 2 months ago)

that’s fine, but no one is saying there aren’t alcohol resistant bacteria, just that the problem you think you’re having & the problem you’re actually having aren’t related. even if the plates are pre poured, if you’re working in the conditions you described there’s still a huge chance for shit to be blown all over the open plate you’re working with. the clear plastic tote SAB with no drafts in the room is what you want :thumbup:


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Offlinematsc
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Re: 70% iso doesn’t kill all bacteria [Re: hazyhorse] * 1
    #28023867 - 10/29/22 11:58 PM (2 years, 2 months ago)

Quote:

hazyhorse said:
that’s fine, but no one is saying there aren’t alcohol resistant bacteria, just that the problem you think you’re having & the problem you’re actually having aren’t related. even if the plates are pre poured, if you’re working in the conditions you described there’s still a huge chance for shit to be blown all over the open plate you’re working with. the clear plastic tote SAB with no drafts in the room is what you want :thumbup:





Definitely agree with ya, but um... I'm not the original poster, just a mildly pedantic forum lurker I'm afraid! Sorry for the confusion :blush:


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OfflineTormatoMFacebookDiscordReddit
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Re: 70% iso doesn’t kill all bacteria [Re: matsc] * 1
    #28023881 - 10/30/22 12:06 AM (2 years, 2 months ago)

I'm just glad nobody mentioned peroxide :uhoh::lol:

:wtf3:


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OfflineGrind365
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Re: 70% iso doesn’t kill all bacteria [Re: Tormato]
    #28023882 - 10/30/22 12:13 AM (2 years, 2 months ago)

Peroxide


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OfflineTormatoMFacebookDiscordReddit
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Re: 70% iso doesn’t kill all bacteria [Re: Grind365]
    #28023890 - 10/30/22 12:27 AM (2 years, 2 months ago)

Quote:

Grind365 said:
Peroxide



You just had to didn't you. :facepalm: :lol:

:shock::aweshit:


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Offlinehazyhorse
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Re: 70% iso doesn’t kill all bacteria [Re: matsc]
    #28023897 - 10/30/22 12:41 AM (2 years, 2 months ago)

Quote:

matsc said:
Definitely agree with ya, but um... I'm not the original poster, just a mildly pedantic forum lurker I'm afraid! Sorry for the confusion :blush:




LMAO ignore me, my bad friend!! my brain sometimes doesn’t register it’s a different person when people don’t have an avi. please continue your pedantry as you see fit

:loveheart:


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OfflineGrind365
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Re: 70% iso doesn’t kill all bacteria [Re: Tormato]
    #28023898 - 10/30/22 12:44 AM (2 years, 2 months ago)

Quote:

Tormato said:
Quote:

Grind365 said:
Peroxide



You just had to didn't you. :facepalm: :lol:

:shock::aweshit:



Sorry. At least I didn't drop the link to the video of the guy touching petri dish after using iso, peroxide and just bare finger.

I may go find the video now though the world needs to know.


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Offlinechris77
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Re: 70% iso doesn’t kill all bacteria [Re: Grind365]
    #28023950 - 10/30/22 02:29 AM (2 years, 2 months ago)

:laugh2:


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OfflineEnvypenis
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Re: 70% iso doesn’t kill all bacteria [Re: chris77]
    #28024021 - 10/30/22 05:42 AM (2 years, 2 months ago)

First off I have to thank you all for quick responses. I tried on Reddit last year, let’s just say this forum blows it away.

I was trying to write that quick before I had to go to bed last night. Work seven days a week no rest for the wicked. I’ll put up some pictures first to clarify then go thru the replies..



“Still air box”.. decent arm holes I thought. Makes a good seal around my arms with lab coat on hence why I got it. I’ll have to pay attention next time and maybe video so you all can see but I really don’t jotice much movement of the material while I’m working. Obviously make a huge reach and there’s a chance. I will pay more attention to speed inside the air box. Read about a few guys going in to do agar work naked. That might be enough to keep me aware of what I’m supposed to be doing in terms of not making quick movements.



This is the air inlet. 6” pvc pipe with correction merv 16 rated filter paper tightened down with hose clamp. Outside is on a carbon filter mounted fan made for weed. The idea was to push positive pressure in the tent so when I would leave a small vent open to let out alcohol fumes it would not drag back in particles. I call the tent my lab, and thought I had a SAB.

The consensus seems to be too much movement within the SAB. I have a 100qt box downstairs I’ll heat cut a couple arm holes tonight, kill all air movement in the tent, and see if it makes any difference. Really can’t thank you all enough on the responses. Flow good is next. Is it generally considered acceptable to work quicker in front of a flow hood? I’m doing 100+ dishes a week and already lacking on time





Hillbilly done in that “SAB”.. I had perfect results in this thing originally. I mean to the point I was confident enough I was pouring 10-15 jello shots at once before putting on covers with near perfect success. I’d Try not to cross over the cups but still happened with better success rates than now. The only other thing that changed was going from pint jars to quart jars but already dealt with that - headspace issue so I thought. Seems to be better now as success rates did improve 30% or so.

Guess I’ll be improving on still air technique vs sterile






Edited by Envypenis (10/30/22 06:07 AM)

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OfflineKROM
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Re: 70% iso doesn’t kill all bacteria [Re: Envypenis] * 1
    #28024036 - 10/30/22 06:16 AM (2 years, 2 months ago)

SAB tek by bod. Making armholes with a coffee can = chef’s kiss.


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OfflineSexBurrito
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Re: 70% iso doesn’t kill all bacteria [Re: Envypenis]
    #28024054 - 10/30/22 06:53 AM (2 years, 2 months ago)

Definitely try a rigid plastic box and kill the fan.  I don't even wear gloves in mine nor do I set it up in my tent anymore and it's works out great, I'd say 95+% of the time.  People would cringe at how I do things but I rarely get contamination. Oddly enough I've also done it naked and didn't really see any difference overall though. Mushrooms and agar work are super easy once you dial in what works for your environment. Your on the right track to succeeding, best of luck


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Re: 70% iso doesn’t kill all bacteria [Re: SexBurrito]
    #28024060 - 10/30/22 07:00 AM (2 years, 2 months ago)

just here to add that you dont want tight cuffs for arm holes. you want wide comfy armholes much bigger than your arms so you can move in and out of them without disturbing the air. i know its counterintuitive but its very well tested


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Re: 70% iso doesn’t kill all bacteria [Re: Baba Yaga]
    #28024063 - 10/30/22 07:04 AM (2 years, 2 months ago)

Quote:

Baba Yaga said:
You have this POS right?



"designed with you in mind"



"hey! do you know nothing about SABs? then we designed this just for you!"


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OfflineSirPsycho
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Re: 70% iso doesn’t kill all bacteria [Re: Baba Yaga]
    #28024073 - 10/30/22 07:13 AM (2 years, 2 months ago)

Quote:

Baba Yaga said:
You have this POS right?




This is the reason for your failure. You will be better off throwing this disgrace of SAB away. I can't believe those ass wipes are actually selling something like this and their advertising is making me throw up, "designed with you in mind", yeah right.

Sorry for the rant it's not your fault, but honestly get rid of this thing and build a SAB out of a rigid plastic tote. Reduce your alcohol use to a minimum and rely on good sterile technique......you know, the kind of technique you weren't able to achieve with this floppy SAB these dickheads sold to you.



Oh fuck me sideways, I'd rather do open air transfers than use that trash bag.... Hell I'll take an actual trash bag over that


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Re: 70% iso doesn’t kill all bacteria [Re: Envypenis]
    #28024165 - 10/30/22 08:40 AM (2 years, 2 months ago)

I never understood why people are putting a SAB inside a tent..I guess it might be useful if you're unable to turn off a fan/air registers in the room, otherwise, think of the SAB as a tiny little tent for your work.

Ditch the flexible SAB or at the very least cut bigger arm holes in it. As others have said you do not want the 'cuffs' to form a seal around your arms. The ONLY thing those SABs have going for them is the clarity. Other than that it's trash.

Quote:

Envypenis said:Read about a few guys going in to do agar work naked.




I'm a nudist and even I can't see a benefit of doing lab work naked


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OfflineDave Bowman
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Re: 70% iso doesn’t kill all bacteria [Re: bigfootscreepyuncl]
    #28024259 - 10/30/22 09:59 AM (2 years, 2 months ago)

Just build a legit SAB made out of a hard plastic.  They are super cheap.

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OfflineEnvypenis
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Re: 70% iso doesn’t kill all bacteria [Re: bigfootscreepyuncl]
    #28024291 - 10/30/22 10:31 AM (2 years, 2 months ago)

The naked part I don’t think would apply to you as a nudist lol As a non nudist I’d be thinking dude you’re naked with every wisp of air that goes by keeping me constantly thinking slow down. The plastic air box I had a year ago then went to open air transfer in the tent, and I guess didn’t put it together that sealed was worse so went with that vinyl set up I think for the size and clarity. I work in my basement so the tent helps me keep a clean area without getting into concrete walls and rafters with dusts/molds everywhere. I also keep my agar in there and temps set to 80*.  Now if the issue is movement of the air box walls.. what if you cut arm holes big enough not to touch the flexible sides at all? And really maintain not touching the sides.. slow down and have deliberate planned and perfectly executed movements within the air box. Is there a write up for still air box work or procedure? Haven’t had time to look while im at work today but need answers when I get home. Quickly replying as I can to keep answers coming

Edited by Envypenis (10/30/22 10:32 AM)

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Re: 70% iso doesn’t kill all bacteria [Re: Envypenis] * 1
    #28024308 - 10/30/22 10:52 AM (2 years, 2 months ago)

logically flexible walls arent ideal for an apparatus with the sole  purpose of staying still, but seeing as you already bought the thing you are welcome to cut wider holes and see if that helps

most people here will correctly point out that a real SAB is so easy and cheap that theres no reason not to get it over with and just fuckin make one. you can keep that other thing around for incubation, use it as a cat condo, or sell it to the next sucker

i like the idea of a tent as a workspace if drafts in the room are hard to control. otherwise its just a placebo-tent

and to whoever suggested using a flow hood in a closed tent would create turbulence...remember that the hood wont even create turbulence at point blank range. that is a key feature of its design


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Re: 70% iso doesn’t kill all bacteria [Re: bakedbeings] * 1
    #28024442 - 10/30/22 12:54 PM (2 years, 2 months ago)

if you have $500 and are in the US/somewhere amazon has the deal, there are FFUs that work as flowhoods. 100000x better than an SAB to work in, would make your work very quick. but if not, the hard plastic SAB is the way to go


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Re: 70% iso doesn’t kill all bacteria [Re: hazyhorse] * 2
    #28024700 - 10/30/22 03:50 PM (2 years, 2 months ago)


Ol' faithful.  Cost like 14$ and works fantastic.  Don't have the money for a flow hood just yet but this just proves that if your method is tight, rudimentary equipment like this will work.  Good luck brother


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Re: 70% iso doesn’t kill all bacteria [Re: SexBurrito]
    #28024806 - 10/30/22 05:08 PM (2 years, 2 months ago)

Really appreciate the help you guys. I knew I was missing something simple but I hate asking for help until I’ve tried everything I can think of. I grew up working in a shop no one had time to answer questions this is just how I handle things now. Still driving home from work right now. I’m going to do a thorough search into still airbox techniques as soon as I’m home but does anyone have anything to add I may not see in the teks?

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Re: 70% iso doesn’t kill all bacteria [Re: hazyhorse]
    #28024852 - 10/30/22 05:34 PM (2 years, 2 months ago)

Inoculate the world had a 2x4 FFU for $1050. That’s my next planned investment. Anyone see anything wrong with that before I make another terrible buy ? Otherwise I’ll go all out and get a lab type set up. Expensive sure.. Had found lab type horizontal flow hoods for around 3300 just a few months back. Can’t seem to find it now. but with all the time I put into this, with lack of time.. seems to be the only option. Plus I’m too stupid to ever quit. Flow hood definitely won’t go to waste

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Re: 70% iso doesn’t kill all bacteria [Re: Envypenis]
    #28024920 - 10/30/22 06:06 PM (2 years, 2 months ago)

ah yes...youre learning. ask before you do things. it took me too long to figure that out

regarding flow hoods i have the YJINGRUI FFU from amazon. i like it a lot, but if i could afford a 2x4 model i would go for it


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Re: 70% iso doesn’t kill all bacteria [Re: bakedbeings] * 1
    #28024949 - 10/30/22 06:15 PM (2 years, 2 months ago)

personally, I'd suggest getting good at a SAB before that kind of investment. For one, if the FFU breaks or you're out of power and need to work you better know how. But also, and probably more important, make sure that you actually enjoy this hobby before dropping that kinda cash on it. That's just my $0.02 - do what you want, I couldn't actually care less :lol:


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Re: 70% iso doesn’t kill all bacteria [Re: bakedbeings] * 1
    #28024950 - 10/30/22 06:16 PM (2 years, 2 months ago)

Always learning .. usually the hard way…

I just read the SAB tech by bod It didn’t take much to click. Really wish I could have seen a year ago. Always have to pay some tuition in the school of hard knocks

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Re: 70% iso doesn’t kill all bacteria [Re: Envypenis]
    #28024958 - 10/30/22 06:18 PM (2 years, 2 months ago)

No worries on that I have a tendency to become obsessed with things and my life will shortly revolve entirely around it.. Practically already does just like to see more come back lol i’m showering now because I leave for work Irish and come home Haitian. I already have another qt of agar warming up in the water bath and will be going down to get my hundred quart tub going to take one of my pots in the kitchen to heat it up about 8 inches and press it through for arm holes and see what the hell happens. I’ll follow the procedure from the tech really take it down to super slow motion. Only change might be i think I’m going to use Lysol to sanitize because they claim 99.9% of germs killled vs alcohol 67% .. Is that going to ruin everything?

Edited by Envypenis (10/30/22 06:22 PM)

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Re: 70% iso doesn’t kill all bacteria [Re: Envypenis]
    #28024964 - 10/30/22 06:20 PM (2 years, 2 months ago)

A flow hood isn't a replacement for sterile technique. Working in front of a flow hood needs just as careful handling as an SAB, the specifics of sterile technique in front of a hood vary quite a bit compared to in an SAB because the two work on almost completely different concepts.


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Re: 70% iso doesn’t kill all bacteria [Re: bigfootscreepyuncl]
    #28024974 - 10/30/22 06:24 PM (2 years, 2 months ago)

Quote:

bigfootscreepyuncl said:
personally, I'd suggest getting good at a SAB before that kind of investment. For one, if the FFU breaks or you're out of power and need to work you better know how. But also, and probably more important, make sure that you actually enjoy this hobby before dropping that kinda cash on it. That's just my $0.02 - do what you want, I couldn't actually care less :lol:



:whathesaid:
totally agree

and also now that im rocking a ffu, im finding that certain things were easier in a sab. stillness is incredibly useful


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Re: 70% iso doesn’t kill all bacteria [Re: SirPsycho]
    #28024979 - 10/30/22 06:27 PM (2 years, 2 months ago)

Correct but I do feel I will be much more comfortable working where I can see and move without being hunched or kneeling.. I hate looking thru the plastic box also. As long as you keep your clean materials upstream of dirtier materials I don’t see much else to go wrong.. sterilization of tools and that’s about it?

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Re: 70% iso doesn’t kill all bacteria [Re: SirPsycho]
    #28024985 - 10/30/22 06:29 PM (2 years, 2 months ago)

Could you expand upon the idea of sterile technique? What did I miss? This is where confusion was found as a beginner because sterile seems to be the goal But apparently is impossible

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Re: 70% iso doesn’t kill all bacteria [Re: bakedbeings]
    #28024992 - 10/30/22 06:31 PM (2 years, 2 months ago)

What In your opinion is easier in the sab vs a flow hood?? That type of info would be great for others to find in the future

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Re: 70% iso doesn’t kill all bacteria [Re: Envypenis] * 1
    #28024994 - 10/30/22 06:31 PM (2 years, 2 months ago)

Quote:

Envypenis said:
As long as you keep your clean materials upstream of dirtier materials I don’t see much else to go wrong.. sterilization of tools and that’s about it?



Yep, it's that simple. Just like SAB work is as simple as not stirring up the nasties.



Quote:

Envypenis said:
move without being hunched or kneeling



Sounds like user error to me. Put your box in a more comfortable spot.


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Re: 70% iso doesn’t kill all bacteria [Re: SirPsycho]
    #28025008 - 10/30/22 06:34 PM (2 years, 2 months ago)

Easy enough to fix. I’m going to take my time building this one now that I have a workplace I call home.. Was a shit show in the beginning.. still a shit show but atleast had a decent harvest last month. Revived my hopes and it’s not just pushing thru hell anymore. Love your quote on balance .. very acccurate!

Edited by Envypenis (10/30/22 06:37 PM)

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Re: 70% iso doesn’t kill all bacteria [Re: Envypenis] * 2
    #28025026 - 10/30/22 06:39 PM (2 years, 2 months ago)

awesome love to see folks not get POd when folks tell them they are doing it wrong!

nope just read about SAB and teks. move slow take your time, dont put hand over anything, I like to rub everthing down JUST incase, give time for things to settle. assume your hand rains down dirt and that everthing is covered in thick uber fine dust that will get stired up with any air movement.

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Re: 70% iso doesn’t kill all bacteria [Re: Envypenis] * 1
    #28025030 - 10/30/22 06:40 PM (2 years, 2 months ago)

Quote:

Envypenis said:
What In your opinion is easier in the sab vs a flow hood?? That type of info would be great for others to find in the future



for me doing small things like agar transfers is more natural with still air but for big movements like pouring LC not being in a box is nice


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Re: 70% iso doesn’t kill all bacteria [Re: the man]
    #28025033 - 10/30/22 06:41 PM (2 years, 2 months ago)

Quote:

the man said:
awesome love to see folks not get POd when folks tell them they are doing it wrong!




:whathesaid:

there are two types of people :/


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Re: 70% iso doesn’t kill all bacteria [Re: Envypenis] * 1
    #28025036 - 10/30/22 06:42 PM (2 years, 2 months ago)

Quote:

Envypenis said:
Could you expand upon the idea of sterile technique? What did I miss? This is where confusion was found as a beginner because sterile seems to be the goal But apparently is impossible



Sure, I absolutely can.

The goal of sterile technique/aseptic work practices is to ensure we don't cause anything floating around to float into our sterile containers.

It's important to understand what is sterile and what is not. The insides of containers that have been pressure cooked are sterile, the outside isn't. The parts of our tools exposed to flame are sterile, the rest isn't. Plates sealed in sleeves from the manufacturer are also sterile.

We don't accomplish our goal of keeping the nasties floating around from getting into our containers by cleaning them away because no matter how hard we try we won't get everything and all it takes is a few spores.

We accomplish our goal through careful handling. In an SAB this pretty much means you never really want to open containers fully, you only lift lids enough to get your tools in there. The times that you do need to completely remove lids, nothing passes over the open containers.

I haven't read your posts in detail but from skimming it seems your problem is the air in the tent is moving and causing the air in your box to move. The room you work in should also be allowed to settle. Any windows, vents, fans should be shut, plugged, turned off.


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Re: 70% iso doesn’t kill all bacteria [Re: the man] * 1
    #28025054 - 10/30/22 06:47 PM (2 years, 2 months ago)

Visualization is a great tool! That will help for sure when I’m down there. Thanks to everyone who chimed in. I mean when you’ve tried everything.. and failed over and over again.. it’s a pretty humbling experience. People who claim ego death on mushrooms.. should try growing it first 🤦♂️

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Re: 70% iso doesn’t kill all bacteria [Re: SirPsycho]
    #28025070 - 10/30/22 06:50 PM (2 years, 2 months ago)

Perfect thank you! For whatever reason I had way too much trust in alcohol to kill the bad stuff. I guess the hand sanitizer push the last few years made it seem more effective. I’m sure some of you guys have scientific backgrounds but I’m assuming there is a pretty big learning curve with this field of study…

I see people using containers like whiskey bottles for pouring which will on the list, smaller mouthed pc safe bottles... But how crazy do I have to be about this. If I lift the lid of the agar, to pour, how could I not get the jar over the dish if pouring into dish below?

Edited by Envypenis (10/30/22 06:53 PM)

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Re: 70% iso doesn’t kill all bacteria [Re: Envypenis]
    #28025087 - 10/30/22 06:56 PM (2 years, 2 months ago)

Quote:

Envypenis said:
Perfect thank you! For whatever reason I had way too much trust in alcohol to kill the bad stuff. I guess the hand sanitizer push the last few years made it seem more effective. I’m sure some of you guys have scientific backgrounds but I’m assuming there is a pretty big learning curve with this field of study..



The key word there is sanitizer. It's not a sterilizer. Sanitary means 99.9% of germs are killed. We need 100%. Only fire and steam/pressure do that, UV too but not for any application that is practical to us really.

The other thing is germs(I.E. shit that has GERMINATED) aren't what we're worried about cause those are easy to kill, it's the spores that haven't germinated that we're worried about. Very few things are sporucidal. Alcohol certainly isn't.


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Re: 70% iso doesn’t kill all bacteria [Re: SirPsycho]
    #28025090 - 10/30/22 06:57 PM (2 years, 2 months ago)

idk bro thats why i stick to no-pours

regarding sterile technique
just imagine that any unsterile surface is covered in contam that can transfer by contact, any air that isnt laminar flow or completely still is full of contam that wants to land in your precious container, and any unsterile object you wave around can shed contam either down (SAB) or towards you (LFH)

you are playing goalie for a sterile food source, against invisible balls


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Re: 70% iso doesn’t kill all bacteria [Re: Envypenis]
    #28025091 - 10/30/22 06:58 PM (2 years, 2 months ago)

Quote:

Envypenis said:
I see people using containers like whiskey bottles for pouring which will on the list, smaller mouthed pc safe bottles... But how crazy do I have to be about this. If I lift the lid of the agar, to pour, how could I not get the jar over the dish if pouring into dish below?



Generally people keep the necks wrapped in foil from the pressure cooker until pouring, keeping the part that will be over plates sterile


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Re: 70% iso doesn’t kill all bacteria [Re: bakedbeings]
    #28025094 - 10/30/22 06:59 PM (2 years, 2 months ago)

Quote:

bakedbeings said:
i stick to no-pours




:whathesaid:

For different reasons though.


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Re: 70% iso doesn’t kill all bacteria [Re: SirPsycho]
    #28025136 - 10/30/22 07:23 PM (2 years, 2 months ago)

Okay that makes sense with the foil over the bottle
Neck. Before coming to shroomery and asking what I was doing wrong.. next option was Pyrex Petri dishes to attempt no pours. What other reasons do you justify the no pour?

Edited by Envypenis (10/30/22 07:23 PM)

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Re: 70% iso doesn’t kill all bacteria [Re: Envypenis] * 1
    #28025157 - 10/30/22 07:34 PM (2 years, 2 months ago)

no pour is nice bc i don’t have to worry about my sterile technique while pouring since they come out of the PC sterile. you have to PC the agar anyways so why not just get it done all at once? also you’re not constantly tossing petri dishes this way. glass petris offer pretty sick visibility but even traditional pasty plates kick ass.


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Re: 70% iso doesn’t kill all bacteria [Re: hazyhorse] * 1
    #28025174 - 10/30/22 07:44 PM (2 years, 2 months ago)

Quote:

hazyhorse said:
no pour is nice bc i don’t have to worry about my sterile technique while pouring since they come out of the PC sterile. you have to PC the agar anyways so why not just get it done all at once? also you’re not constantly tossing petri dishes this way. glass petris offer pretty sick visibility but even traditional pasty plates kick ass.



:rockon:


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Re: 70% iso doesn’t kill all bacteria [Re: bakedbeings]
    #28025188 - 10/30/22 07:55 PM (2 years, 2 months ago)

Definitely going in the no pour direction when i can. I do love being able to see what I’m working with. It just makes sense anyways. One less possible chance of contamination. I’ll get back to you all by tomorrow night with results. Praying I’ll see an improvement. Going to be extremely anal about this batch

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Re: 70% iso doesn’t kill all bacteria [Re: Envypenis]
    #28025191 - 10/30/22 07:58 PM (2 years, 2 months ago)

Personally I go with pre-pours because I only have a 6qt pressure cooker, and it would drive me bonkers trying to fit a bottle of media in there...

But I can fit 10 'plates' at a time in it just fine, which is more than I really need at any given moment anyway.

The condensation is a little annoying, I will admit, but nothing too awful.


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Re: 70% iso doesn’t kill all bacteria [Re: matsc]
    #28025210 - 10/30/22 08:16 PM (2 years, 2 months ago)

Never had any problems with pouring in a SAB but I prefer no pours cause it's something that is more suitable to do alongside other stuff.

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Re: 70% iso doesn’t kill all bacteria [Re: Envypenis]
    #28025225 - 10/30/22 08:31 PM (2 years, 2 months ago)

Quote:

Envypenis said:
Okay that makes sense with the foil over the bottle
Neck. Before coming to shroomery and asking what I was doing wrong.. next option was Pyrex Petri dishes to attempt no pours. What other reasons do you justify the no pour?



I basically don't have a permanent place for my SAB, it gets put away after each session. I don't want to deal with timing the set up, worrying about temp, then having to let them cool


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Re: 70% iso doesn’t kill all bacteria [Re: SirPsycho] * 2
    #28026466 - 10/31/22 04:23 PM (2 years, 2 months ago)

Alright so after digging into a few decent reads on this site.. I decided to ditch just about everything I “learned.” After reading just a few basic beginners threads here I noticed I made the ultimate mistake of YouTube and Reddit. I’m sure you guys must love Willy myco..

I need clarification on whether agar can caramelize if over cooked or re sterilized? Quick google search tells me sugars don’t start to Carmelize until 336* .. I’ve found some contradicting answers on this one.

I ditched the quart jars I had made up the night before, and re bottled them. I had quart media jars in my cabinet the whole time. Loosened cover before PC and covered with foil. 40 mins pressure and just a CH above zero I released pressure and as quickly as possible Sealed the media bottles. Will get 500 ml soon didn’t like having to tip them over in the PC but it worked.



I made up my 100 qt rigid still air box with nice roomy 8” arm holes. Very comfortable now I guess the 66 qt was my downfall Before. I have the agar in the water bath now at 120*. Have a few more questions before I start tonight..

Towel or no towel? The edges of the tub don’t sit flat on the table. 1/8” gap in areas. Towel would make an even seal but again I don’t know if that’s better or worse.

I’m out of Petri dishes and have to resort to using the Jell-O shot cups. Going to make it interesting. Thinking I’ll lay out the cups with lids resting on top one by one then go ahead and pour. Can easily see the advantages here now of the dishes.

I can also see why the tent would be worse for SAB work. I swear if I breathe I can see the walls move a minute amount never mind moving my body in this bitch. I’ll be very much paying attention this round. Appreciate all the constructive criticism and eye openers.. seriously woke me the fuck up

Edited by Envypenis (10/31/22 04:25 PM)

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Offlinehazyhorse
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Re: 70% iso doesn’t kill all bacteria [Re: Envypenis]
    #28026481 - 10/31/22 04:32 PM (2 years, 2 months ago)

glad we could help man! thanks for taking the advice to heart. i think it was a good idea to get rid of all your prior “knowledge”

i’ve never had problems resterilizing agar a few times, you should be fine

i’ve seen people on here recommend the towel on the bottom. i wouldn’t work directly on the towel obviously, but if it’s gonna seal your SAB better on the bottom i think it’s a good idea. i’ve heard it helps contams from stirring up (as long as you’re not moving the towel around or something obviously)


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Re: 70% iso doesn’t kill all bacteria [Re: hazyhorse] * 1
    #28026491 - 10/31/22 04:37 PM (2 years, 2 months ago)

the damp towel is a classic trick, i assume a dry towel would work. get yourself a nice big rack that fits the sab. like an oven rack or cooling rack for bakes goods, something like that

dont worry about agar carmelizing


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Re: 70% iso doesn’t kill all bacteria [Re: hazyhorse] * 1
    #28026498 - 10/31/22 04:39 PM (2 years, 2 months ago)

Thank you this place really is amazing. What I read last night I’m sold. You all are changing the world with this at home science. It’s incredible. Loved the part about we basically are all committing felonies don’t get butthurt over simple words! I understand it all. Especially squashing bad info.. to make sure good info persists. I’m sure I’ll be a life long member now

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Re: 70% iso doesn’t kill all bacteria [Re: Envypenis]
    #28026504 - 10/31/22 04:41 PM (2 years, 2 months ago)

you’re gonna fit in great around here. excellent attitude to have my friend. welcome to the shroomery :cheers:


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Re: 70% iso doesn’t kill all bacteria [Re: Envypenis]
    #28026506 - 10/31/22 04:41 PM (2 years, 2 months ago)

Quote:

Envypenis said:


I made up my 100 qt rigid still air box with nice roomy 8” arm holes.





That is a nice SAB with well sized arm holes :thumbup:


What does the lid of this tub look like? Some can be used as SAB floors if they are flat and without any weird grooves and such nonsens.

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Re: 70% iso doesn’t kill all bacteria [Re: bakedbeings]
    #28026513 - 10/31/22 04:45 PM (2 years, 2 months ago)

Damn always one more thing.. plastic bags hung over the arm holes. Do you guys tape them on and remove when you go to work?  roll them up out of the way? Tape from the side and fold off to the side when working? I don’t think I’ll have to worry about this tonight as the Jell-O shots have a snap top. But when I do get Petris again you hang the bags while waiting for the dishes to cool before parafilm correct?

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Re: 70% iso doesn’t kill all bacteria [Re: Baba Yaga]
    #28026515 - 10/31/22 04:46 PM (2 years, 2 months ago)

Unfortunately it’s a pain in the ass cover. Ridges and a huge bow in the middle somehow. I really couldn’t get the rack to sit flat without it being dead center taking up most of the space

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Re: 70% iso doesn’t kill all bacteria [Re: Envypenis]
    #28026521 - 10/31/22 04:51 PM (2 years, 2 months ago)

tbh i just covered my armholes by draping a t shirt  off the top & slightly pushed the tshirt in the holes. there is probably a more elegant solution but it worked for me


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Re: 70% iso doesn’t kill all bacteria [Re: hazyhorse]
    #28026531 - 10/31/22 04:59 PM (2 years, 2 months ago)

yea same i drape a big towel over it. i imagine plastic covers taped on would be a pain to deal with


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Re: 70% iso doesn’t kill all bacteria [Re: bakedbeings]
    #28026732 - 10/31/22 06:52 PM (2 years, 2 months ago)

Perfect even more simple.. just finished pouring 1.6L of agar. I tried one bottle no towel, one bottle with towel. I can see how the mist would also help to catch the contam so why not try both on the stainless table. I’m definitely still in the experimental phase. Gave me a chance to wipe everything out again and start fresh in between batches. Very much looking forward to Petri dishes but I don’t think it went horribly with Jell-O shots. Only thing I didn’t really like was I Couldn’t press the containers closed on the rack. I had to move it down to the cutting board to press closed. But all was covered before moving down to contamination land. Honestly I was at 10-30% success rate before so I’d say anything will be better

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Re: 70% iso doesn’t kill all bacteria [Re: hazyhorse]
    #28026742 - 10/31/22 07:03 PM (2 years, 2 months ago)

What’s your opinion on flipping agar dishes? Always flip? Only flip after pouring? Or flip when you have inoculated with agar wedge also? And if you flip with agar wedge how long would you let it set to adhere to the plate? need to dig a little deeper myself into this topic but have found contradicting answers again I believe. I was flipping agar cups with agar wedges and they kept falling onto the lid.. Never really could find an answer how long you let them adhere but seemed like overnight worked.. 

Edited by Envypenis (10/31/22 07:05 PM)

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Re: 70% iso doesn’t kill all bacteria [Re: Envypenis]
    #28026748 - 10/31/22 07:06 PM (2 years, 2 months ago)

I've never once flipped a plate. I don't think that is common practice in this hobby


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Re: 70% iso doesn’t kill all bacteria [Re: bigfootscreepyuncl]
    #28026755 - 10/31/22 07:09 PM (2 years, 2 months ago)

Quote:

bigfootscreepyuncl said:
I've never once flipped a plate. I don't think that is common practice in this hobby



Maybe 20 years ago


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Re: 70% iso doesn’t kill all bacteria [Re: bigfootscreepyuncl]
    #28026760 - 10/31/22 07:10 PM (2 years, 2 months ago)

Awesome thank you that was what I pretty much got when I did look into it. For bacterial cultures you would flip, not for mycology. Definitely helps ease the mind

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Re: 70% iso doesn’t kill all bacteria [Re: Envypenis]
    #28026776 - 10/31/22 07:18 PM (2 years, 2 months ago)

Quote:

Envypenis said:
Awesome thank you that was what I pretty much got when I did look into it. For bacterial cultures you would flip, not for mycology. Definitely helps ease the mind



I don't know about growing bacteria, I hear it's quite challenging to do it properly. For our purposes though there's really no need to flip. The water in your plates should be just as sterile as the surface of the agar.

I put plates with condensation at the bottom of the stack and if I have the opportunity before they dry I'll use them for spore germination


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Re: 70% iso doesn’t kill all bacteria [Re: SirPsycho]
    #28026783 - 10/31/22 07:22 PM (2 years, 2 months ago)

That’s a helpful tip I’ll go rotate the condensated cups to the bottom right now. My first batch I was at 127* when I went down and had condensation. The second batch was 117* and not a drop of condensation on any. I’ll definitely be replicating that from here on out

Edited by Envypenis (10/31/22 07:23 PM)

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Re: 70% iso doesn’t kill all bacteria [Re: Envypenis]
    #28026796 - 10/31/22 07:28 PM (2 years, 2 months ago)

Quote:

Envypenis said:
Awesome thank you that was what I pretty much got when I did look into it. For bacterial cultures you would flip, not for mycology. Definitely helps ease the mind



I have and never will flip a plate upside down for any reason, especially if it's got condensation on the lid. All yours gonna do is flip it and dump the water onto the plate.

Just be careful when you move them around and removing the lid when you use em.

There's ways to get rid of it, but I'm lazy as fuck and I just don't worry about it. :lol:

I feel the condensation helps keep the plate fresh longer. If there's condensation they can't be dried out, right? :shrug: :lol:


That's just how I get down. :awegroove:


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Re: 70% iso doesn’t kill all bacteria [Re: bakedbeings]
    #28026866 - 10/31/22 08:15 PM (2 years, 2 months ago)

Quote:

bakedbeings said:
idk bro thats why i stick to no-pours

regarding sterile technique
just imagine that any unsterile surface is covered in contam that can transfer by contact, any air that isnt laminar flow or completely still is full of contam that wants to land in your precious container, and any unsterile object you wave around can shed contam either down (SAB) or towards you (LFH)

you are playing goalie for a sterile food source, against invisible balls




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Re: 70% iso doesn’t kill all bacteria [Re: Envypenis]
    #28026884 - 10/31/22 08:22 PM (2 years, 2 months ago)

I use wash cloths and binder clips.  Washcloths lightly damp with soapy water or bleach water clipped to the top of the arm hole and let to drape down.

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Re: 70% iso doesn’t kill all bacteria [Re: SymPlayTon] * 1
    #28027106 - 10/31/22 10:30 PM (2 years, 2 months ago)

Nice changes Envy

sure that new SAB is gonna serve you well

all the best with it going forward :rockon:

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Re: 70% iso doesn’t kill all bacteria [Re: Tormato]
    #28027546 - 11/01/22 08:36 AM (2 years, 2 months ago)

I can’t believe how much I overcomplicated this whole deal. One less waste of time lol I had noticed If I had condensated cups and older agars that had dried out a bit.. take the drier wedge to the wetter agar and would sometimes come back for me. Are there any other tricks for trying to save dried agar samples? The last few batches of agar I effed up royally I really needed those genetics to continue on. Most have dried too much by now.

Preliminary report and probably way too early to tell.. but ALL agar cups from last night using properly adjusted technique looked clean this morning. Have had the tent set at 82* and will hopefully have same results in another 12 hrs

Agar was setting pretty good by the end of pouring and some cups didn’t set entirely flat. Some maybe a little bumpy on the surface. Only thing I couldn’t tell what it was.. could see some shiny spots but no cloudiness what so ever. Not sure if that was condensate that fell and got absorbed?

Edited by Envypenis (11/01/22 08:44 AM)

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OfflineTheUsualSuspect
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Re: 70% iso doesn’t kill all bacteria [Re: Envypenis]
    #28028712 - 11/01/22 09:31 PM (2 years, 2 months ago)

Quote:

Envypenis said:
I can’t believe how much I overcomplicated this whole deal.




I assure you, everyone does that. My first grow I did spores to grain bag and had a nasa looking mini tent with humidifyer and fans and sensors... etc.

Needless to say the second attempt looked completely different.

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