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Offlinemake air go wooo
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Registered: 12/02/21
Posts: 60
Last seen: 10 months, 1 day
sanitation questions
    #27960320 - 09/21/22 08:35 AM (1 year, 4 months ago)

I apologize if some of these questions sound stupid. My equipment is a SAB with kitchen cooling racks lining the bottom.

1) When using a disposable inoculation loop, do you wipe down the wrapper with alcohol outside of the SAB?
(1A) What do you normally do with the wrapper?

2) If you have to set the inoculation loop down between inoculating petri dishes, where do you put it?
(2A) Do you ever sanitize the disposable inoculation loop after opening it?

3) When using a store bought pre-sterilized blade to cut out mycelium, do you have to sterilize the part of the handle that reaches over the top of the petri dishes?
(3A) If you have to set the scalpel down between work, where do you put it?

4) How do you store your used blades to prevent rust?

5) Is it best to wipe things down with alcohol inside the SAB?
(5A) Are crappy paper towels okay to use? I fear they leave lint.

6) Is it ok to use the plastic petri dishes that come in sleeves of 20, to squirt some spores into? It's from a spore syringe for the purpose of dipping your inoculation loop.
(6A) Do you spray the petri dish wrapper with alcohol before you put it into the SAB?
(6B) Do you need to wipe the inside or outside of the petri dishes after taking them out of wrapper, inside the SAB?

My goal here is to develop better sanitation and sterilization procedures.
Again, I apologize for the mundayne questions, but these are personalized questions. Thank you Shroom folk.


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OnlineKmacmo
The aborted pin
Other


Registered: 08/14/19
Posts: 1,675
Loc: Central hemisphere
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Re: sanitation questions [Re: make air go wooo]
    #27960475 - 09/21/22 10:39 AM (1 year, 4 months ago)

1. Yes wipe outside then put inside SAB
1a. Keep wrapper inside SAB remove it when your done.

2. I use a empty petri dish as a mini table inside the SAB.
2a. If it's pre-steralized then you don't need to.

3. Yes. The handle should be wiped with ISO before its put inside SAB.
3a. I put it on my mini petri dish table.

4. I bin all my used blades.

5. All wiping with ISO should be done outside SAB.
5a. I've had no problems with cheap kitchen paper.

6. That's a good idea.
6a. Yes anything you put inside the SAB should be ISO cleaned before it goes in.

6b. No. They should be pre-steralized petri dishes no need to wipe the dish only wipe the wrapper.


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Offlinemake air go wooo
Stranger

Registered: 12/02/21
Posts: 60
Last seen: 10 months, 1 day
Re: sanitation questions [Re: Kmacmo]
    #27960513 - 09/21/22 11:03 AM (1 year, 4 months ago)

I thought of another question.

If I wanted to pour some agar plates, and leave them sit to see if they grow mold, without being inoculated, can I just let them sit inside the SAB with NO parafilm? How many days max would you let them sit in the SAB?

So far, from experience, the agar I leave unwrapped grows mold.

Thanks for your response


Edited by make air go wooo (09/21/22 11:13 AM)


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Invisiblebakedbeings
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Registered: 09/01/20
Posts: 4,218
Re: sanitation questions [Re: Kmacmo] * 1
    #27960530 - 09/21/22 11:17 AM (1 year, 4 months ago)

try to find a steady place to lean tools while you arent working, so the business end is in the air. sometimes i make a lil taco out of foil, but you can use a glass/jar/whatever

i use 1 blade for everything and i just keep it in a drawer like you would store a regular knife. i flame it before using it anyway so it doesnt matter what it touches. rust isnt a concern imo

idk why kmacmo says wiping must be done outside the SAB. for some things its easier but i generally put everything in my SAB and let the air settle before wiping it all down. many ways to skin these cats i suppose

if youre inoculating with a syringe why not just skip the loop and put a drop on each plate? loops make more sense with prints imo

leaving test plates unsealed in your SAB is valid if you seal the holes to keep the air still. if youre going to be walking around those open holes creating air currents it defeats the purpose


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OnlineKmacmo
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Registered: 08/14/19
Posts: 1,675
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Re: sanitation questions [Re: bakedbeings]
    #27960594 - 09/21/22 12:10 PM (1 year, 4 months ago)

Quote:

Bakedbeings said:
idk why kmacmo says wiping must be done outside the SAB. for some things its easier but i generally put everything in my SAB and let the air settle before wiping it all down. many ways to skin these cats i suppose




Your right there's different ways of doing it. Glad you commented I was under the impression that cleaning inside the SAB might risk contaminating the inside or the air inside the SAB?


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Invisiblebakedbeings
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Re: sanitation questions [Re: Kmacmo] * 1
    #27960651 - 09/21/22 12:51 PM (1 year, 4 months ago)

*shrugs* it works for me


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OfflineSirPsycho
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Re: sanitation questions [Re: bakedbeings] * 2
    #27960720 - 09/21/22 01:33 PM (1 year, 4 months ago)

You don't really need to wipe the outside of anything, I certainly don't. As for where you put your tools while they aren't being used that doesn't really matter either, you should be flaming them before every transfer (if you're doing multiple pieces of agar into one jar you only need to flame when you move to the next jar)

Spraying alcohol or anything really while working is definitely a no no, you're creating movement in a STILL AIR box.

Good aseptic work practices in an SAB are not about cleaning, they are about careful handling such that the only things that touch the inside of or hover above your sterile containers are flame sterilized tools.

This involves careful hand placement so you don't put your hands over open containers, carefully grasping lids so your fingers don't roll over inside edges, only lifting lids enough to get your tools in there, not putting the un-sterilized portions of your tools in the containers.


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:pm:Ask me about free Ps tampanesis, Ps subtropicalis and Ps cubensis (ESS) prints:pm:
Balance in life is like running on ice.

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Offlinemake air go wooo
Stranger

Registered: 12/02/21
Posts: 60
Last seen: 10 months, 1 day
Re: sanitation questions [Re: SirPsycho]
    #27960947 - 09/21/22 04:15 PM (1 year, 4 months ago)

What if your inoculation multiple petri dishes. Do you flame sterilize in-between each one?


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OfflineSirPsycho
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Registered: 01/01/20
Posts: 6,900
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Trusted Cultivator
Re: sanitation questions [Re: make air go wooo]
    #27960970 - 09/21/22 04:24 PM (1 year, 4 months ago)

Yes, every time you switch containers. Donor or receiver


--------------------
:pm:Ask me about free Ps tampanesis, Ps subtropicalis and Ps cubensis (ESS) prints:pm:
Balance in life is like running on ice.

  🅑🅞🅣🅣🅛🅔 🅖🅐🅝🅖

"Mist your balls and fan your asshole" - Pandaskis, 2023


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OfflinePBJ710
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Registered: 07/05/19
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Re: sanitation questions [Re: SirPsycho] * 1
    #27961949 - 09/22/22 05:26 AM (1 year, 4 months ago)

While disposable loops work well, you will go through hundreds or thousands of them if you try to use them appropriately.  Thats why everyone uses a metal loop that can be heat sterilized.  It doesn't have to be pretty - a stiff bread tie or some scrap wire will work.

I'm still using the same #11 blade that I started the year with.  Just wipe the handle/heat the blade between each culture.  When the tip starts curling over from being heated too much I'll swap it out with a fresh one.


Edited by PBJ710 (09/22/22 05:33 AM)


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