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Benson
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Registered: 09/29/20
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How long after pouring agar do I have to wait before inoculation?
#27116169 - 12/30/20 12:19 PM (3 years, 29 days ago) |
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I've read through a bunch of teks and tried searching for this but I haven't found anything . On D3monic's agar tek he said:
Quote:
Once all plates are poured leave unwrapped in SAB with arm holes covered for several days or several hours in front of flowhood to allow condensation to subside.
But I want to inoculate sooner than a few days. What's the consensus?
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coversall
إِنْ شَاءَ ٱللَهُ



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Re: How long after pouring agar do I have to wait before inoculation? [Re: Benson] 1
#27116173 - 12/30/20 12:21 PM (3 years, 29 days ago) |
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If you're confident in your technique then go for it. If you're aren't than waiting some days to check the plates are clean is a good way to save spores. A stich in time saves nine.
Plus allowing the condensation to subside will be helpful in the long run.
Why the rush?
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Land Trout
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Re: How long after pouring agar do I have to wait before inoculation? [Re: coversall] 1
#27116192 - 12/30/20 12:30 PM (3 years, 29 days ago) |
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I wait until they cool so maybe an hour, or more. It works.
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Benson
The Kidd ⭐


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Re: How long after pouring agar do I have to wait before inoculation? [Re: coversall]
#27116194 - 12/30/20 12:30 PM (3 years, 29 days ago) |
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Hmm, good points. I can only go into my lab once every few days for the next month or so; just trying to be efficient with my time while getting setup!
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Benson
The Kidd ⭐


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Re: How long after pouring agar do I have to wait before inoculation? [Re: Land Trout]
#27116198 - 12/30/20 12:33 PM (3 years, 29 days ago) |
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Quote:
Land Trout said: I wait until they cool so maybe an hour, or more. It works.
Nice I think I'll pour a bunch and try half soon, half later. Thanks bois
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PBJ710
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Re: How long after pouring agar do I have to wait before inoculation? [Re: coversall] 1
#27116201 - 12/30/20 12:33 PM (3 years, 29 days ago) |
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You CAN inoculate as soon as it's cool enough that it won't harm the mycelium. As coversall said, the wait is verify the cleanwork was done correctly and let condensation evaporate. If you're confident that you're not causing any contam issues on the plates and that the recipe is good, go for it.
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Land Trout
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Re: How long after pouring agar do I have to wait before inoculation? [Re: PBJ710]
#27121829 - 01/02/21 10:47 AM (3 years, 26 days ago) |
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Thanks for posting this question. It’s helping me dial in my plates. Always thought getting cultures on them sooner was better, but I’m letting some sit, and liking the results.
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Smelltheflowers
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Re: How long after pouring agar do I have to wait before inoculation? [Re: Land Trout]
#27130170 - 01/06/21 11:33 AM (3 years, 22 days ago) |
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I’m new to agar and cultivating mushrooms. After pouring agar plates should I wrap with parafilm while waiting? And what is a good rule of thumb on how long to wait to put liquid culture on new agar plates?
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mushhead
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Re: How long after pouring agar do I have to wait before inoculation? [Re: Smelltheflowers]
#27130176 - 01/06/21 11:35 AM (3 years, 22 days ago) |
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If you're sterilization procedure is on spot and they've cooled down and set proper? Use em. wrap up what you're not going to use first. That is AFTER they set and cooled. It takes less than an hour for it all to be set and ready to use for me. About 45min. I do stacks of 10 per stack in my SAB, usually do about 100+ plates at a time. I use parafilm to wrap my plates, hope you've had practice with it, it does take a bit of time to "wrap" your head around it.
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Smelltheflowers
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Re: How long after pouring agar do I have to wait before inoculation? [Re: mushhead]
#27130212 - 01/06/21 11:54 AM (3 years, 22 days ago) |
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Mush head, this will be my very first time. I’ve been reading as much as possible about it all before diving in. I really want to get it right. I have a question about the SAB and sterilization. Is it best to use 70% alcohol to wipe the outside and inside of SAB? Or 50 water/50bleach? Also, is it ok to put a soaked towel of 70; alcohol on the bottom of SAB and a metal rack on top of soaked alcohol towel to work on? Of course without using lighter or flammables. I just want to make sure I don’t contam anything.
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mushhead
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Re: How long after pouring agar do I have to wait before inoculation? [Re: Smelltheflowers]
#27130235 - 01/06/21 12:05 PM (3 years, 22 days ago) |
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Quote:
Smelltheflowers said: Mush head, this will be my very first time. I’ve been reading as much as possible about it all before diving in. I really want to get it right. I have a question about the SAB and sterilization. Is it best to use 70% alcohol to wipe the outside and inside of SAB? Or 50 water/50bleach? Also, is it ok to put a soaked towel of 70; alcohol on the bottom of SAB and a metal rack on top of soaked alcohol towel to work on? Of course without using lighter or flammables. I just want to make sure I don’t contam anything.
Here's my process. I scrub out my SAB using hot soapy water. I then clean out my SAB with iso and microban (lysol spray) Bleach is corrosive, dangerous, and will burn off your nose hairs. So I don't recommend using it at all, but in a pinch it will work just fine. I would just put down a rack that sits about 3inches or more above the bottom of your SAB. Next is all just sterile technique. You will need to use a torch or alcohol lamp to sterilize you tools used within the SAB. Make your scalpels, needles, innoc loops, and anything else metal used for cult get red hot and allow to cool a little before use.
Edited by mushhead (01/06/21 12:31 PM)
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Marxcelium
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Re: How long after pouring agar do I have to wait before inoculation? [Re: mushhead] 1
#27130261 - 01/06/21 12:18 PM (3 years, 22 days ago) |
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I'll add that, instead of wrapping plates that haven't been inoculated, you can squeeze five at a time into a sandwich ziplock (manufacturing process leaves the inside of the bags sterile) or just put them all back in the bag they came in if you remove them carefully enough to keep the inside of the bag clean.
For my time, its a pain in the ass to wrap and unwrap a bunch of plate you haven't even inoculated yet.
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mushhead
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Re: How long after pouring agar do I have to wait before inoculation? [Re: Marxcelium]
#27130312 - 01/06/21 12:36 PM (3 years, 22 days ago) |
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Yep most ziplock bags are sterile before you open them. So if you open them within your SAB to store your plates within them that'll work too. However you should keep the plates upright so they don't go falling over and getting contamed.
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Smelltheflowers
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Re: How long after pouring agar do I have to wait before inoculation? [Re: mushhead]
#27130324 - 01/06/21 12:40 PM (3 years, 22 days ago) |
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Mush head, do you let the alcohol in the SAB settle a while before bringing in the plates and agar bottle? Or bring them in right after and let it settle? Then wipe with alcohol inside the SAB?
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mushhead
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Re: How long after pouring agar do I have to wait before inoculation? [Re: Smelltheflowers]
#27130354 - 01/06/21 12:50 PM (3 years, 22 days ago) |
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I just wipe out my SAB of everything as much as I can, sometimes there some liquid left but otherwise my SAB is mostly dry when I begin using it. EDIT: I will post a picture of my SAB and set up so you can see everything I'm working with. Please note that it will be sanitized and ready to use in the picture, which will give you a great starting point to designing your own set up.
Edited by mushhead (01/06/21 01:17 PM)
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Marxcelium
Mushroom Instrumentality Project

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Re: How long after pouring agar do I have to wait before inoculation? [Re: Smelltheflowers]
#27130474 - 01/06/21 01:26 PM (3 years, 22 days ago) |
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Just a heads up from experience, I'd be incredibly careful with spraying alcohol in your SAB and then bringing flamed tools into it. The SAB will trap the alcohol vapor as it dries and then ignite if the tool is still hot enough.
Soapy water is all I used, and it works great. I liked to leave the walls and floor of the SAB under my rack wet with soap water to trap contams so my movements didn't stir them back into the air.
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mushhead
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Re: How long after pouring agar do I have to wait before inoculation? [Re: Marxcelium] 1
#27130499 - 01/06/21 01:33 PM (3 years, 22 days ago) |
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Quote:
Marxcelium said: Just a heads up from experience, I'd be incredibly careful with spraying alcohol in your SAB and then bringing flamed tools into it. The SAB will trap the alcohol vapor as it dries and then ignite if the tool is still hot enough.
Soapy water is all I used, and it works great. I liked to leave the walls and floor of the SAB under my rack wet with soap water to trap contams so my movements didn't stir them back into the air.
if he's using 70% there no worries. You'd have to be worried if it were 99.5% iso. I use an open cup of iso to dip my tool in inside my SAB, even put hot scalpels into it without any flames at all ever. Not once in my years of using iso inside my SAB, on my SAB, or near my SAB has caused any kind of explosion or flames other than what I intend which is my torch. If you like I'll post video of the whole process I go through. iso is nothing to be worried over exploding unless you're being way fucking stupid.
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Marxcelium
Mushroom Instrumentality Project

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Re: How long after pouring agar do I have to wait before inoculation? [Re: mushhead] 1
#27130520 - 01/06/21 01:40 PM (3 years, 22 days ago) |
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Just trying to raise a little stupid guy awareness then in case there are any dumb fuckers like me out there
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mushhead
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Re: How long after pouring agar do I have to wait before inoculation? [Re: Marxcelium]
#27130605 - 01/06/21 02:09 PM (3 years, 22 days ago) |
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WASH OUT YOUR SAB WITH HOT SOAPY WATER BEFORE APPLYING ANY KIND OF CHEMICAL TREATMENT, SOAP BINDS TO DIRT AND OTHER PARTICLES. STERILE TECHNIQUE IS JUST AS IMPORTANT AS CLENLINESS
 Here is everything I use to clean out my SAB. Going from left to right the first two are emergency use only when I can't find iso or lysol/microban. I do not recommend you use bleach at all ever unless forced. Then we have lab grade 99.5% alcohol which I then make 72% iso out of which is next to it and finally the lysol spray I like. Plus alcohol wipes.
 These are my tools, this consists of parafilm, media bottles for agar/LC, plastic gloves/heat resistant gloves, Temp gun, and box full of various needle tips, scalpel blades, agar, etc it's an everything box.
 This is the vessel I have inside my SAB which is the 72% alcohol. I dip my scalpel in this after EVERY transfer made ever without fail, then flame sterilize it. The combination I find really helps with keeping your scalpels clean. I don't recommend this for needles though just fyi.
 This is inside my SAB. I have a rack here which sits about 2 1/2inches off the bottom of it. I also have a cup to hold my scalpels, syringes, inocc loops when they're not in use, and of course my alcohol cup for assisting with sterilization.
 Enjoy. M.H
Edited by mushhead (01/06/21 10:34 PM)
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Smelltheflowers
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Re: How long after pouring agar do I have to wait before inoculation? [Re: Marxcelium]
#27130609 - 01/06/21 02:10 PM (3 years, 22 days ago) |
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Mush head and Marxcelium, thanks for the input. I seriously want to be mindful about this. I definitely do not want any dangerous accidents! Mush head please post the video you use when you get a chance. It will be much appreciated. I’m going to go with a soapy towel at the bottom of my SAB to be on the safe side 😁
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