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OfflineShrooms4fun
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Some thoughts
    #23970286 - 12/31/16 09:45 AM (7 years, 1 month ago)

So I have been playing around the last several months with different things such as agar and g2g transfers and here and there i seem to keep getting contamination  (trichoderma), I ended up with 2 of 12 jars after a g2g being contaminated and the petri dishes appear to be really really prone to contamination. I'm doing all of my work inside a SAB that I spray out with 70% ISO and let sit for a bit while wearing a complete tyvek suit also with the sleeves sprayed off with ISO. Now we all know how expensive a flow hood is either to build or buy, it seems you can't really get out of investing anything less than about $300 into one. So here is my thought for a solution to this issue seeing as how a flow hood is a hepha filter I would like to place a small desktop hepha filter inside the SAB to improve the sterility inside my work environment such as something like this: Desktop hepha filter

Furthermore and beyond the air filtration within my SAB this hepha also talks about UV light sterilization of the air, I have e briefly read threw a few things talking about UV serialization and how it is not suitable for things like substrates.....but how about for things like plastic petri dishes or syringes, without the needle as its not transparent and can just be ran threw the PC anyhow. Would something like UV light be suitable for sterilization in those applications?

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Invisibler.lutece
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Re: Some thoughts [Re: Shrooms4fun]
    #23970293 - 12/31/16 09:49 AM (7 years, 1 month ago)

The point of a still air box is just that it has little to no moving air. Introducing a HEPA into it implies there will also be air blowing into it, which defeats the purpose of an SAB.

As far as the UV sterilization goes, I fear that you'd be hard-pressed to find any effective UV sterilizer sold as part of a desktop HEPA filter.


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One goes into an experiment knowing one might fail.
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Offlineenlightenment
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Re: Some thoughts [Re: Shrooms4fun]
    #23970301 - 12/31/16 09:53 AM (7 years, 1 month ago)

It is possible to get great results with a SAB even if your environment is polluted. How do you prepare the agar? BTW Penicillium and Aspergillus look similar to trichoderma for an unexperienced eye.
The SAB might not be the problem. It could be the way you prepare your agar and grain.

//edit

Have you tried to make your SAB work in another small room? Closed doors/windows.

Edited by enlightenment (12/31/16 10:01 AM)

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OfflineShrooms4fun
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Re: Some thoughts [Re: enlightenment]
    #23970324 - 12/31/16 10:01 AM (7 years, 1 month ago)

Quote:

enlightenment said:
It is possible to get great results with a SAB even if your environment is polluted. How do you prepare the agar? Penicillium and Aspergillus look similar to trichoderma for an unexperienced eye. So the SAB might not be the problem. It could be the way you prepare your agar and grain.




Prepoured dishes from one of the sponsors here at the shroomery meay dishes  the g2g jars I'm sure I did something wrong with as well because all donations and recipient jars sat for quite some time uncontaminated.....perhaps moving to quickly?

Yes I am aware that the point t of a still air box is for the air to remain  still and isolate the outside contaminants that was why I had brought the he question up because it seems to be the opposite of what I am proposing....However a flowhood is constant movement in front of your work as well that was filtered so that's where this idea came to mind.

If you read the hehe description of the hehe he link above the hehe heat filter says it has a UV light inside it as part of the filtration system....I just wanted to take that a step further and buy for example a UV light to sterilize plastics and such kinda like how they erase eeproms with a UV light

Edited by Shrooms4fun (12/31/16 10:59 AM)

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InvisiblebodhisattaMDiscordReddit
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Re: Some thoughts [Re: Shrooms4fun]
    #23970346 - 12/31/16 10:10 AM (7 years, 1 month ago)

sounds like you just need to develop SAB skills. No lights or filter will make up for lack of technique

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Offlinemrmazdarx9
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Re: Some thoughts [Re: Shrooms4fun]
    #23970413 - 12/31/16 10:38 AM (7 years, 1 month ago)

Quote:

korn16ftl3 said:
Quote:

enlightenment said:
It is possible to get great results with a SAB even if your environment is polluted. How do you prepare the agar? Penicillium and Aspergillus look similar to trichoderma for an unexperienced eye. So the SAB might not be the problem. It could be the way you prepare your agar and grain.




Prepoured dishes from one of the sponsors here at the shroomery meay dishes  the g2g jars I'm sure I did something wrong with as well because all donations and recipient jars sat for quite some time uncondemned.....perhaps moving to quickly?

Yes I am aware that the point t of a still air box is for the air to remain  still and isolate the outside contaminants that was why I had brought the he question up because it seems to be the opposite of what I am proposing....However a flowhood is constant movement in front of your work as well that was filtered so that's where this idea came to mind.

If you read the hehe description of the hehe he link above the hehe heat filter says it has a UV light inside it as part of the hehe he ha the filtration system....I just wanted to take that a step further and buy for example a UV light to sterilize plastics and such kinda like how they erase eeproms with a UV light



Whats with all the hehe hehe hehe is your keyboard/pad broke or something?
Also buying expensive filtration systems and uv lights wont make up for what is clearly poor technique


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InvisiblebodhisattaMDiscordReddit
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Re: Some thoughts [Re: mrmazdarx9]
    #23970421 - 12/31/16 10:41 AM (7 years, 1 month ago)


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OfflineShrooms4fun
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Re: Some thoughts [Re: bodhisatta]
    #23970456 - 12/31/16 10:55 AM (7 years, 1 month ago)

No its the autocorrect and the damn phone guessing what I'm going to type crap....it's driving me insane I gotta find a way to turn it off sorry about that I will edit the reply correctly


So it seems to be here that people are voting it as tenique flaws.....how do I use process of elimination to sort out exactly where my flaws are at? I can see using smooth slow continous motions as a potential flaw of mine....ive never been that great at it.

Edited by Shrooms4fun (12/31/16 11:06 AM)

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Offlinemrmazdarx9
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Re: Some thoughts [Re: Shrooms4fun]
    #23970535 - 12/31/16 11:27 AM (7 years, 1 month ago)

Also how large are your arm holes have you got a pic? Are you spraying the inside with soapy water before you start?


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OfflineShrooms4fun
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Re: Some thoughts [Re: mrmazdarx9]
    #23973559 - 01/01/17 05:05 PM (7 years, 1 month ago)

Quote:

mrmazdarx9 said:
Also how large are your arm holes have you got a pic? Are you spraying the inside with soapy water before you start?



7" in diameter is the hole size for my SAB in a really large tote (I can get the measurements and/or take a picture of it tomorrow as its not at my home location. As far as spraying the insides of the SAB I use 70% ISO alchol no soapy water.

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Offlineenlightenment
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Re: Some thoughts [Re: Shrooms4fun]
    #23973575 - 01/01/17 05:11 PM (7 years, 1 month ago)

I think you already know it but anyway:
Be careful with fire next to your SAB and never try to flame something inside the SAB if you sprayed it with 70% iso.

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Invisibler.lutece
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Re: Some thoughts [Re: Shrooms4fun]
    #23973758 - 01/01/17 06:07 PM (7 years, 1 month ago)

Quote:

korn16ftl3 said:
Quote:

mrmazdarx9 said:
Also how large are your arm holes have you got a pic? Are you spraying the inside with soapy water before you start?



7" in diameter is the hole size for my SAB in a really large tote (I can get the measurements and/or take a picture of it tomorrow as its not at my home location. As far as spraying the insides of the SAB I use 70% ISO alchol no soapy water.




Soapy water is actually preferable to iso for spraying the walls before you start working. Iso is great for wiping surfaces that can't be flame sterilized and need to be interacted with directly (gloved hands, injection ports, grain jars, etc.), but when you spray the walls, you're not looking to sanitize them. It's actually to create a wet, semi-sticky surface so that any contams that drift in will hopefully get stuck to them. Iso dries too quickly for this purpose, but water sticks around a little longer. Adding a little soap helps to prevent the water from evaporating as quickly.


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OfflineTh3Issu3
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Re: Some thoughts [Re: r.lutece]
    #23973884 - 01/01/17 06:48 PM (7 years, 1 month ago)

What do your jar lids look like?

how many transfers are you doing from your plates ? what do they look like ?

How long are you PCing your grain ? Are you letting it steam up for 5-10 mins then placing the rocker on ?

LOT of different variables here.

I went months getting contams when it was my lids not properly done.

Edited by Th3Issu3 (01/01/17 06:51 PM)

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OfflineShrooms4fun
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Re: Some thoughts [Re: r.lutece]
    #23974251 - 01/01/17 09:20 PM (7 years, 1 month ago)

Ok so I'll change that part of my process then and see how things vary.

Now would a UV light box be sufficient to sterilize clear things like petri dishes and syringe cylinders?

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InvisibleAdden
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Re: Some thoughts [Re: Shrooms4fun] * 1
    #23974267 - 01/01/17 09:27 PM (7 years, 1 month ago)

Why are you spraying your SAB with iso?
Load a spray bottle with a little bit of antibacterial soap. Clean off a table with soapy water, discard towel. Lay down a fresh towel and spray it until damp. Flip your SAB over and spray the 5 sides. Wipe it clean. Spray again with soapy water. Flip it again onto the towel. Sometimes I'll spray a few more spritzes of soapy water when it's proper, just cuz you lose some to evap so fast.

Lay your stuff out and load in your 70% iso'd materials, let sit 15 minutes and go to work. You don't have to iso your gloves and stuff either. Just don't get your hands over any media. My arm holes are 7 inches cuz I got big arms doesn't seem to make any difference. Be as methodical and robotic as you can with smooth, calculated movements.

If you get new tools or new jar sizes or whatever, do a dry run with no media and see how everything stacks up. Then replicate those movements when you have all your gear and mats together.

Edited by Adden (01/01/17 10:03 PM)

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InvisibleTookitooki
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Re: Some thoughts [Re: Adden] * 1
    #23974294 - 01/01/17 09:37 PM (7 years, 1 month ago)

:whatshesaid:

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