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JHOVA
Post whore


Registered: 02/17/17
Posts: 4,727
Loc:
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Re: Pastywhyte's Easy Agar Tek [Re: Hobbit GDF]
#26253518 - 10/15/19 10:20 AM (4 years, 3 months ago) |
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Do nothing
-------------------- ๐
๐ด ๐ฐ ๐ผ ๐ฒ ๐ป ๐ธ ๐ฝ ๐ถ ๐
๐
๐ฐ ๐ฟ
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pixelpopper
Crap Artist

Registered: 09/20/13
Posts: 4,022
Loc: Dreamland
Last seen: 3 months, 11 days
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Re: Pastywhyte's Easy Agar Tek [Re: JHOVA]
#26253523 - 10/15/19 10:21 AM (4 years, 3 months ago) |
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Either let them sit long enough for the water to absorb, or what I do sometimes is open them sideways and then carefully dump the water out onto ISO papertowel inside of SAB
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spirit_shadow
Feature not a bug



Registered: 08/15/11
Posts: 25,672
Last seen: 1 hour, 20 minutes
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Re: Pastywhyte's Easy Agar Tek [Re: pixelpopper]
#26259676 - 10/17/19 08:57 PM (4 years, 3 months ago) |
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Just posting to ask something: did they discontinue the mini rounds? I have checked every local store in a few towns radius and none have them. I had to order mine online :/
-------------------- ERROR 418 IM A TEAPOT.....(this account is automated, all posts related to illegal activities or advice thereof are strictly from numerous online sites and are for informational purposes only)- Circa 2011 Ban lotto
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Northern Thailand
Stranger

Registered: 08/29/19
Posts: 13
Last seen: 4 years, 1 month
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Have I missed it?!? How much water goes in the PC?
TIA, Charlie
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McDominator



Registered: 08/29/19
Posts: 270
Loc: California
Last seen: 1 year, 2 months
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Hey Guys, so I tried this tek a couple of times now, and both times I got contams growing even though I did nothing with the agar. Wondering if someone can spot check my process.
So this last time around I made them in these screw top zip lock containers:


Here is my pressure cooker, which I believe only gets up to 11.6 psi or so:

Here was the result after PCing for 1.5 hours and letting them cool in the PC..... for a week. As recommended to avoid condensation.

I am using 2 layers of 3m paper micropore tape as my filter. This time around everything seemed soaked when I took it out, paper towel, tape, everything. I'm assuming that is not a good thing. They were wrapped in foil. I put just a small layer of water in the bottom of the pot when I PCed, maybe like 1.5 cups.
Right now I'm thinking it's either my PC isn't up to par being only 11psi, or something to do with this damn tape.
It seems to be the same sour smelling white bacteria build up every time. I'm just glad I didn't waste spores this time around!
Help a newb out? 
EDIT: Doing some research I found this. Seems like micropore tape is suspect:
https://www.shroomery.org/forums/showflat.php/Number/13642707
Edited by McDominator (10/18/19 10:25 AM)
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primelines
Agarth Brooks


Registered: 02/25/16
Posts: 484
Last seen: 5 days, 12 hours
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What kind of water are you using? City? Well water? Try distilled maybe.
Is your GE hole in the side of the container? It should be on the top.
Fuck that pc get a presto.
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the_sonic16
Mushy McMusherson



Registered: 03/10/19
Posts: 396
Loc: Somewhere off topic
Last seen: 2 years, 18 days
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It could be that you cooked them for too long. I've been reading posts around here trying to calibrate the appropriate agar cook time for an Instant Pot and the best I came up with was 45 minutes. I did that and mine came out looking fine. (As to whether or not they're sterile, I'll have to let you know ).
Anyway, figured I'd throw my hypothesis out there. We'll probably need to work through this one collaboratively, as there are a growing number of IP users and not a comprehensive amount of documentation surrounding how it can be used for mycology.
Also, regarding the GE hole on the side, there was an update to the tek later that suggested that as an option so as to eliminate the need for papertowels and foil.
-------------------- Sonic's "I'll buy a pressure cooker someday" Instant Pot times and settings:
- Popping Corn Rapid Re-hydration - Half fill all containers with corn. Empty into Instant Pot. Cover with water. Run 30 minutes on PC. Release, remove, dry.
- Wheat/Rye Berry Soak Re-hydration - Half fill all containers with grain. Empty in pot with water. Soak for 6-12 hours, air dry until surface is completely dry.
- Grain Jar Sterilization - Run 2 - 2.5 hrs on PC
- PastyPlates, MEA Formulation- Run 45 minutes on PC
- LME Liquid Culture - Run 45 minutes on PC
Edited by the_sonic16 (11/04/19 11:28 AM)
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McDominator



Registered: 08/29/19
Posts: 270
Loc: California
Last seen: 1 year, 2 months
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Yea, seriously looking into getting a Presto right now.
And thought I read something about GE hole on the side is okay.
I think I'm going to get a presto and forego the micropore tape for syringe filters.
-------------------- I'm here to learn. I'm also willing to help. If I'm wrong on something, please call me out. I am not resistant to new information, but I always carry a healthy dose of skepticism. โIt would be possible to describe everything scientifically, but it would make no sense; it would be without meaning, as if you described a Beethoven symphony as a variation of wave pressure.โ -Albert Einstein
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CapMeh
RX Jazzist



Registered: 06/08/09
Posts: 924
Loc: The Holy Mountain
Last seen: 2 months, 28 days
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Quote:
McDominator said: Hey Guys, so I tried this tek a couple of times now, and both times I got contams growing even though I did nothing with the agar. Wondering if someone can spot check my process.
So this last time around I made them in these screw top zip lock containers:


Here is my pressure cooker, which I believe only gets up to 11.6 psi or so:

Here was the result after PCing for 1.5 hours and letting them cool in the PC..... for a week. As recommended to avoid condensation.

I am using 2 layers of 3m paper micropore tape as my filter. This time around everything seemed soaked when I took it out, paper towel, tape, everything. I'm assuming that is not a good thing. They were wrapped in foil. I put just a small layer of water in the bottom of the pot when I PCed, maybe like 1.5 cups.
Right now I'm thinking it's either my PC isn't up to par being only 11psi, or something to do with this damn tape.
It seems to be the same sour smelling white bacteria build up every time. I'm just glad I didn't waste spores this time around!
Help a newb out? 
EDIT: Doing some research I found this. Seems like micropore tape is suspect:
https://www.shroomery.org/forums/showflat.php/Number/13642707
1st get a legit PC
2nd don't modify twist top agar dishes. Leave the lids slightly unscrewed and they wont collapse during the PC (Wrap them in foil and unscrew them a quarter turn before putting them in the PC... I'm not even sure if foil is necessary). Then simply re-tighten them when taking them out of the PC or in the SAB after taking them out.
3rd DONT WORRY ABOUT CONDENSATION! Leaving them in a PC for a week to minimize condensation is ridiculous. I use them a day after pressure cooking, many have condensation, and I've never had an issue. I've only used screw top agar dishes with 100% success.
https://www.shroomery.org/forums/showflat.php/Number/26189684
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spirit_shadow
Feature not a bug



Registered: 08/15/11
Posts: 25,672
Last seen: 1 hour, 20 minutes
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Re: Pasty Agar Tek [Re: CapMeh]
#26261989 - 10/18/19 08:47 PM (4 years, 3 months ago) |
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...... Before I got my mini rounds I legit went to dollar store got cheap as fuck tiny plastic containers and tested them with no contams 
Edit: in fact.....
-------------------- ERROR 418 IM A TEAPOT.....(this account is automated, all posts related to illegal activities or advice thereof are strictly from numerous online sites and are for informational purposes only)- Circa 2011 Ban lotto
Edited by spirit_shadow (10/18/19 08:54 PM)
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McDominator



Registered: 08/29/19
Posts: 270
Loc: California
Last seen: 1 year, 2 months
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Re: Pasty Agar Tek [Re: CapMeh]
#26262359 - 10/19/19 04:19 AM (4 years, 3 months ago) |
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Quote:
CapMeh said:
Quote:
McDominator said: Hey Guys, so I tried this tek a couple of times now, and both times I got contams growing even though I did nothing with the agar. Wondering if someone can spot check my process.
So this last time around I made them in these screw top zip lock containers:


Here is my pressure cooker, which I believe only gets up to 11.6 psi or so:

Here was the result after PCing for 1.5 hours and letting them cool in the PC..... for a week. As recommended to avoid condensation.

I am using 2 layers of 3m paper micropore tape as my filter. This time around everything seemed soaked when I took it out, paper towel, tape, everything. I'm assuming that is not a good thing. They were wrapped in foil. I put just a small layer of water in the bottom of the pot when I PCed, maybe like 1.5 cups.
Right now I'm thinking it's either my PC isn't up to par being only 11psi, or something to do with this damn tape.
It seems to be the same sour smelling white bacteria build up every time. I'm just glad I didn't waste spores this time around!
Help a newb out? 
EDIT: Doing some research I found this. Seems like micropore tape is suspect:
https://www.shroomery.org/forums/showflat.php/Number/13642707
1st get a legit PC
2nd don't modify twist top agar dishes. Leave the lids slightly unscrewed and they wont collapse during the PC (Wrap them in foil and unscrew them a quarter turn before putting them in the PC... I'm not even sure if foil is necessary). Then simply re-tighten them when taking them out of the PC or in the SAB after taking them out.
3rd DONT WORRY ABOUT CONDENSATION! Leaving them in a PC for a week to minimize condensation is ridiculous. I use them a day after pressure cooking, many have condensation, and I've never had an issue. I've only used screw top agar dishes with 100% success.
https://www.shroomery.org/forums/showflat.php/Number/26189684
Yes! I love this. Not needing to modify the container would save a lot of time and cash I was going to spend on syringe filters. Do your cultures still grow fine with no air exchange?
In the write up you did those containers look huge. Wondering if they just contain enough air inside the container because of their size. What size are they?
Definitely going to get a legit PC when I get some funds.
And the only reason I left for a week is because I was waiting on spores. Honestly glad I was forced to wait to see my process was garbage before I ended up wasting spores.
-------------------- I'm here to learn. I'm also willing to help. If I'm wrong on something, please call me out. I am not resistant to new information, but I always carry a healthy dose of skepticism. โIt would be possible to describe everything scientifically, but it would make no sense; it would be without meaning, as if you described a Beethoven symphony as a variation of wave pressure.โ -Albert Einstein
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Drboomer
The lord magnificent



Registered: 09/22/19
Posts: 957
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So I couldn't source the agar bars. About how much agar powder a tsp?
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the_sonic16
Mushy McMusherson



Registered: 03/10/19
Posts: 396
Loc: Somewhere off topic
Last seen: 2 years, 18 days
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Re: Pasty Agar Tek [Re: Drboomer]
#26262947 - 10/19/19 11:30 AM (4 years, 3 months ago) |
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Quote:
Drboomer said: So I couldn't source the agar bars. About how much agar powder a tsp?
Quote:
I then spoon 2 heaping tablespoons into the pot. I rarely weigh it out, if I want stiffer agar I use a little more, if I want softer I use a little less. If you want to weigh it, for this amount of agar you will want about 2-2.5 grams depending on how soft or firm you want it to be. If your using powdered agar and don't have a scale, a level tsp (5ml) is roughly 2.5 grams
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CapMeh
RX Jazzist



Registered: 06/08/09
Posts: 924
Loc: The Holy Mountain
Last seen: 2 months, 28 days
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The big ones are 2 cups. I also use 1/2 cup containers, both seem to work fine. I prefer the 2 cup containers because it gives the mycelium more space to run which is good if I'm letting them fully colonize to use as spawn. Also, my plates will fruit if they fully colonize and consolidate so having extra room can be nice if Iโm busy.
From what Iโve read, and Iโm still a novice, if you want to do agar transfers for the purpose of strain/genetic isolation you should use the 1/2 cup containers. This is because you arenโt letting the agar fully colonize so youโre saving agar in the long run.
Donโt overthink it too much
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the_sonic16
Mushy McMusherson



Registered: 03/10/19
Posts: 396
Loc: Somewhere off topic
Last seen: 2 years, 18 days
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Quote:
McDominator said: Hey Guys, so I tried this tek a couple of times now, and both times I got contams growing even though I did nothing with the agar. Wondering if someone can spot check my process.
So this last time around I made them in these screw top zip lock containers:


Here is my pressure cooker, which I believe only gets up to 11.6 psi or so:

Here was the result after PCing for 1.5 hours and letting them cool in the PC..... for a week. As recommended to avoid condensation.

I am using 2 layers of 3m paper micropore tape as my filter. This time around everything seemed soaked when I took it out, paper towel, tape, everything. I'm assuming that is not a good thing. They were wrapped in foil. I put just a small layer of water in the bottom of the pot when I PCed, maybe like 1.5 cups.
Right now I'm thinking it's either my PC isn't up to par being only 11psi, or something to do with this damn tape.
It seems to be the same sour smelling white bacteria build up every time. I'm just glad I didn't waste spores this time around!
Help a newb out? 
EDIT: Doing some research I found this. Seems like micropore tape is suspect:
https://www.shroomery.org/forums/showflat.php/Number/13642707
I'm not a TC, but I figured I'd take a shot at this since I'm using my IP for everything else. I was able to make MEA Pasty Plates using my Instant Pot.
I ran it on "Pressure Cook", "High Pressure" for 45 minutes. No carmelization and they set just fine. The only thing I need to work on improving is the condensation.
It's been a week and I've grown mycelium on inoculated plates. Conversely, nothing has grown on my control plates. Obviously, that's a short period of time to declare a complete success. I'll post if they suddenly contaminate, but figured I'd share that this is possible in the meantime.
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Shroomhog
Stranger
Registered: 10/02/19
Posts: 2
Last seen: 3 years, 10 months
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Can you tell me how to go from syringe to sterile swab? do i just spray the sterile swab with the syringe spores, then swab on the agar?
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mushboy
modboy



Registered: 04/24/05
Posts: 32,275
Loc: where?
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That's what I do. Wet the swab with a few drops from the syringe and swipe away
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Drboomer
The lord magnificent



Registered: 09/22/19
Posts: 957
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Re: Pasty Agar Tek [Re: mushboy]
#26285258 - 10/29/19 04:05 PM (4 years, 2 months ago) |
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Took my first shot at this, dropped some ms on plates. Was psyched when I got growth 5 days in. It turned green. Remaking now might pc some cotton swabs as sterile swabs arent available here locally. I'll try my inoculation loop too and see how it goes
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Mr. Alien
I will abduct andprobe your anus



Registered: 01/14/14
Posts: 6,290
Loc: Star Wars Galaxy
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Re: Pasty Agar Tek [Re: Drboomer]
#26285998 - 10/29/19 09:56 PM (4 years, 2 months ago) |
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I was thinking using PC sterilized cotton tips, a few dry and a few wet. Then, taking the dry one to rub the spore print to pick up some spore then use the wet tip to "steal" some spores from the dry one, then inoculate with the wet tip the agar plate. This should work i think?
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Northern Thailand
Stranger

Registered: 08/29/19
Posts: 13
Last seen: 4 years, 1 month
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Hi folks, I don't mean to be overly stupid, but I read Pasty's tek four times and I don't see how much water to put in the PC. Am I somehow blinding myself?!
How much water to put in the PC when nuking the rounds?
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