Paradox
©
Fisana

Jump to content


Photo
* * * * * 1 votes

Looking for agar transfer advice (timing, contams)?


  • Please log in to reply
15 replies to this topic

#1 scofos83

scofos83

    Mycophiliac

  • Free Member
  • 12 posts

Posted 17 March 2020 - 10:45 PM

I've been wanting to start growing again after a few PF Teks in my early 20's. The gist is that I wanted to try stepping up a bit and like many others, love tech & automation and have seen the arguments both for and against automation. Long story short, I want to dabble and work towards a proof of concept. So far I obtained spores from Mushrooms(dot)com, was supposed to get 1 PE syringe and 1 pan cyan jamaican print, however they were out of that print so without asking to specify they just sent me the syringe of the cyans too. Ive been reading and watching everything I could on the steps to a next level grow and was able to build a pretty badass (IMO) portable flow hood that uses an inline duct fan to get the CFM to 100 (actual CFM 107).  I created 4 32qt tubs for fruiting and modified a Crane Drop humidifier to handle all 4 tubs with the use of "t's", although I don't expect it to run all the time.
 
To start my flow hood was set up in the doorway of a closet to minimize the air flow around the environment. After spraying and wiping it all down, I used the syringes with pre-poured PDA agar dishes I sourced from Etsy. Next time I'd like to pour my own but I wanted to eliminate at least 1 contam vector and cut my startup work a tiny bit with the plan to make half Penis Envy and half Pan Cyan Jamaican. As for my noob agar procedure lol, I did just one ms drop off of an inoculating loop I made from 18ga SS vape wire that I twisted with a drill and secured in an Exacto knife handle and sterilized. I've never done agar and I def think I need practice so I don't gouge it lol. I dont know if the loop was too big, wire too thin, agar too soft, but when I tried to streak the various lines and patterns it would catch and dig in to the agar. Nonetheless, I flamed the tip in between each streak.
 
Currently for incubation I have all the dishes inside a smaller tub on top of a dry towel, then placed inside  one of the 32qt tubs on top of a seedling mat thats controlled by an Inkbird temp/humid controller keeping it within 5 degrees of 83. Currently there aren't plans for the Inkbird hygrometer; I imagine I could hook a fan up to it but again, I doubt on the small scale it'd be needed much anyway. They have been incubating for 11 days and now im HAPPY to say, I have lots of growth, both good and bad, and likely need to transfer but need to make sure im not grabbing anything im not supposed to lol. The green colonies are obvious, but want to be sure the others are mycelium before doing all the work.  I thought the pan cyans weren't working but then I spotted the thinner and almost transparent look of it as it starts. Some plates still have absolutely nothing still. Any suggestions or feedback regarding the transfers is greatly appreciated. Im excited and grateful to be a part of this community ;-p
 

Attached Thumbnails

  • IMG_20200317_001511.jpg
  • IMG_20200317_001336.jpg
  • IMG_20200317_001032.jpg
  • IMG_20200317_000923.jpg
  • IMG_20200316_234446.jpg
  • IMG_20200316_235205.jpg
  • IMG_20200316_235300.jpg
  • IMG_20200316_235340.jpg
  • IMG_20200316_235408.jpg
  • IMG_20200316_235425.jpg
  • IMG_20200316_235436.jpg
  • IMG_20200316_235514.jpg
  • IMG_20200316_235614.jpg
  • IMG_20200316_235626.jpg
  • IMG_20200316_235642.jpg
  • IMG_20200316_235705.jpg
  • IMG_20200316_235723.jpg

Edited by scofos83, 17 March 2020 - 10:46 PM.


#2 CatsAndBats

CatsAndBats

    This motherfucker

  • OG VIP
  • 12,221 posts

Awards Bar:

Posted 18 March 2020 - 11:07 AM

Here's our cope thread (pan cyan):

 

https://mycotopia.ne...naeolus-thread/

 

There's a wealth of info there. It's pretty much agreed upon that wispy cope myc is superior to tomentose myc. So your cope Jamaican plates are good.

 

Here's a great thread too, addressing some of the taxonomy/etymology of copelandia:

 

https://mycotopia.ne...9-from-florida/

 

Here's more great threads by one of my beloved mentors, peacefrog:

 

https://mycotopia.ne...-pans-grow-log/

 

https://mycotopia.ne...le-and-recipes/

 

Here are some threads by some motherfucker that you might find interesting:

 

https://mycotopia.ne...us-of-my-penis/

 

https://mycotopia.ne...izes-agar-agar/

 

 

What was your agar recipe? Also if one is working from syringe to agar, one can use a sterile swab to streak the syringe liquid on the agar surface, FYI.


  • ChocolateStarfish likes this

#3 scofos83

scofos83

    Mycophiliac

  • Free Member
  • 12 posts

Posted 20 March 2020 - 12:07 AM

Here's our cope thread (pan cyan):

 

https://mycotopia.ne...naeolus-thread/

 

There's a wealth of info there. It's pretty much agreed upon that wispy cope myc is superior to tomentose myc. So your cope Jamaican plates are good.

 

Here's a great thread too, addressing some of the taxonomy/etymology of copelandia:

 

https://mycotopia.ne...9-from-florida/

 

Here's more great threads by one of my beloved mentors, peacefrog:

 

https://mycotopia.ne...-pans-grow-log/

 

https://mycotopia.ne...le-and-recipes/

 

Here are some threads by some motherfucker that you might find interesting:

 

https://mycotopia.ne...us-of-my-penis/

 

https://mycotopia.ne...izes-agar-agar/

 

 

What was your agar recipe? Also if one is working from syringe to agar, one can use a sterile swab to streak the syringe liquid on the agar surface, FYI.

I dont have an agar recipe yet, I bought them as I mentioned from here: https://www.etsy.com...le-petri-dishes Ive heard of the "D" in PDA agar however referred to as both Dextrose Agar and Dung Agar. Which is correct?


  • CatsAndBats likes this

#4 CatsAndBats

CatsAndBats

    This motherfucker

  • OG VIP
  • 12,221 posts

Awards Bar:

Posted 20 March 2020 - 12:12 AM

Dextrose.

 

PDA and MEA are the most common agar recipes (potato dextrose agar and malt extract agar, respectively).


Edited by CatsAndBats, 20 March 2020 - 12:15 AM.


#5 scofos83

scofos83

    Mycophiliac

  • Free Member
  • 12 posts

Posted 20 March 2020 - 03:04 PM

So I checked again today and decided to pull the dishes that still have no growth. Some of these look really nice, but admittedly im still a noob. From what little I know, the single clean circular dishes are great to keep, the mold one looks to have good growth sector to use, but what about the streak dishes with multiple spots? I think there is clean growth to get out of it but should I bother since I have the other better dishes? On the image of 5 plates, most look thin with the exception of the lower 2 and not in the wispy "good" way. Does my assesment sound somewhat accurate? Also, on the same 5 plate image the lower right PE dish seems to be yellowing slightly. I know that can be a lot of things, so my question is how do you rule out Aspergillus etc if it all can look just like mycelium lol?

 

Attached Thumbnails

  • IMG_20200320_153654.jpg
  • IMG_20200320_153722.jpg
  • IMG_20200320_153821.jpg

  • CatsAndBats likes this

#6 CatsAndBats

CatsAndBats

    This motherfucker

  • OG VIP
  • 12,221 posts

Awards Bar:

Posted 20 March 2020 - 03:40 PM

I'd toss the moldy one, it's already sporulated, there's a bazillion spores all over that plate that you can't see.



#7 scofos83

scofos83

    Mycophiliac

  • Free Member
  • 12 posts

Posted 22 March 2020 - 07:02 PM

I'd toss the moldy one, it's already sporulated, there's a bazillion spores all over that plate that you can't see.

Aww man really? I was hoping I could still pick from that one nice edge. When was the tipping point on that one ya think? Suppose ideally should've done right away, but when the other "edge" colonies popped up what that the big tell-tale sign? IM glad i checked, im about to do them in a few minutes so thanks.



#8 CatsAndBats

CatsAndBats

    This motherfucker

  • OG VIP
  • 12,221 posts

Awards Bar:

Posted 22 March 2020 - 11:27 PM

 

I'd toss the moldy one, it's already sporulated, there's a bazillion spores all over that plate that you can't see.

Aww man really? I was hoping I could still pick from that one nice edge. When was the tipping point on that one ya think? Suppose ideally should've done right away, but when the other "edge" colonies popped up what that the big tell-tale sign? IM glad i checked, im about to do them in a few minutes so thanks.

 

Once you see green or black spores (or any color) on an agar plate, the whole plate is compromised.


  • Arathu likes this

#9 scofos83

scofos83

    Mycophiliac

  • Free Member
  • 12 posts

Posted 23 March 2020 - 12:10 AM

 

 

I'd toss the moldy one, it's already sporulated, there's a bazillion spores all over that plate that you can't see.

Aww man really? I was hoping I could still pick from that one nice edge. When was the tipping point on that one ya think? Suppose ideally should've done right away, but when the other "edge" colonies popped up what that the big tell-tale sign? IM glad i checked, im about to do them in a few minutes so thanks.

 

Once you see green or black spores (or any color) on an agar plate, the whole plate is compromised.

 

 

Gotcha. So it's more advised to try and transfer it away from other healthy mycelium for isolation purposes, and not so much in my case with more of a shotgun approach to my syringe? (if that made sense lol) I think I just read enough to confuse myself. :biggrin:



#10 Boebs

Boebs

    Psychonaut

  • VIP
  • 1,165 posts

Awards Bar:

Posted 23 March 2020 - 06:53 AM

If you have enough plates, however you could save that myc in the contaminated plate, however.. it may take a few transfers, and id do it well away from my clean room.
If you have more spores, id restart myself tho.

#11 scofos83

scofos83

    Mycophiliac

  • Free Member
  • 12 posts

Posted 04 April 2020 - 05:10 PM

Checking in, havent been able to update much with all the craziness. Hope the Mycotopia fam is safe and well. I took your advice, and tossed the bad plate. I picked a couple to transfer, some are getting pretty decent I think but Im still struggling with the jamaican pans. I tried a few more plates out of the syringe, this time with a swab but there all slow fat donuts lol. When  would it be ideal to move one to LC?

 

Attached Thumbnails

  • IMG_20200329_210234.jpg


#12 scofos83

scofos83

    Mycophiliac

  • Free Member
  • 12 posts

Posted 11 April 2020 - 06:27 PM

Back with updates, hope everyone is good during amidst all this craziness:) I took some advice on the transfers and while I had everything set up I tried some more pan spores to agar via swab since I don't seem to be getting the fast wispy growth. I've lowered the temps to max 80. I knew I wanted to get to play with LC so with the extra time I took the House of Hydro fan I never used and made a magnetic stirrer. For the rye inoculations, I used sterile syringes to make myc water for half of them, the other half were with agar wedges. I wanted to see the difference for myself:) They have been going for 11 days and seem good.  In one of the pics that I circled twice, would you always want to take a horizontal section or is there a benefit to vertical sections? For the second pic with a singular  section circled, when a section is this defined, would you simply pluck that to transfer? Plus, the dish labeled ISO has a very strong rhizo section I want to use but Im not 100% which way to come at it. I also have my bulk substrate ready to go which I culled from a few sources like JOC and Asura which was manure, straw, vermiculite, and a lil gypsum, pc'ed and cooled. I dont understand why the media uploader here is so bad, so I screenshot the images lol.
 
 

Attached Thumbnails

  • Screen Shot on 2020-04-11 at 19:26:29.png

  • Arathu likes this

#13 Arathu

Arathu

    Dirtmaker

  • OG VIP
  • 7,035 posts

Awards Bar:

Posted 15 April 2020 - 06:53 PM

post-113856-0-93279800-1586993282.jpg

 

If you can see where I put the red circles on the two plates (row 2,col 2 and row 2, col 3), personally I'd be taking transfers just about the size of those circles, maybe smaller if I can see it, getting the leading edge of those running, nay charging rhizomes....expand something like that and see where it takes you.....

 

Next time you have a plate or two with something similar I'd say go for it. Obviously you have the idea of what to do with it afterwards. Work on the plates can be quite fruitful later.....

Cats had a, Malabar was it?, that was crazy aggressive and rhizome madness.

 

Yep it's right here.....

post-147940-0-20018500-1478049070.jpg

 

From this thread https://mycotopia.ne...-agar-30/page-8

 

Well worth the read and study if you haven't been there yet.....I need to go back and see all the goodies I missed there in fact......

 

Nice bags.....I really like filter patch bags too...

 

A

Attached Thumbnails

  • Untitled.jpg

Edited by Arathu, 15 April 2020 - 06:54 PM.

  • Tenderfoot, CatsAndBats and YoshiTrainer like this

#14 CatsAndBats

CatsAndBats

    This motherfucker

  • OG VIP
  • 12,221 posts

Awards Bar:

Posted 16 April 2020 - 10:01 AM

I saw that pic and recognized it immediately. :biggrin:


  • Arathu likes this

#15 scofos83

scofos83

    Mycophiliac

  • Free Member
  • 12 posts

Posted 23 April 2020 - 11:05 PM

So definitely getting what I asked for with learning and dialing new auto setups in lol :laugh: I had spawned to bulk sub of manure/straw, verm, and some gypsum. Cased with peat/verm/calcium carbonate. Misted and placed back in original closet. Due to drastic temp flux I made the decision (with girls approval haha) to move into the closet in the other room. After wiping down and cleaning I set it up the same which was 2 tubs stacked on each side with the humidifier between them, along with a heating pad both hooked to Inkbird controller. Everything seemed good once set up, however got case of noob paranoia coupled with the threads reminding over and over you cant just crank it up and walk away so I kept checking every hour or so and went to bed. I woke up in the am to inkbird alarm on humidity because excessive condensation had pooled in the tube leading to the control tub and blocked the humidity and air flow.  Im guessing lack of FAE, high humidity/temp, and/or a rushed setup introduced me to cobweb mold. And wow it IS impressively fast. The one tub went from small dime size to 3/4 of casing surface overnight and a second has small circular splotch.  I treated with Hydrogen peroxide spray and seemed to clear it up. I fixed the condensation issue by using a space heater in the actual room and tweaked the humidity system based on JOC tek idea with central reservoir.  I also got a Germ guardian air purifier that had $50 rebate:), UV, and Hepa to clean and add some flow. I first had it up on the top shelf on its side angled to blow downward towards the closet door and a small Vornado personal size fan to add some flow back up.
 
Ive since scrapped that too and changed setup which is 2 tubs on the right, 1 on left (ultimately woke to the tub totally covered in light gray cobweb so I tossed it), I added a new tub I got on the floor on top of my seedling mat to bump up ambient humidity, put the purifier on top still blowing a door (around 150cfm may be much), I put the heating pad on the shelf with a 4" AC Infinity duct fan on top with few inches of duct hose on the front and back. Essentially pushing air down past tubs to back of purifier, towards door, and back up again. Since making these changes I saw huge difference in temp and rh stability. Ive been able to hold around 75-80 deg @ 90-97% RH with what appears to be great evap rate, possibly too good? Its very humid in there but there is no vapor droplets on walls and sub surface "looks" dry, but not sure. Im nervous to keep humidity on surface high because I dont want that particular tub to succumb to the cobweb again. Today I see some growth on this PE tub in question, but hard to say if its cobweb or mycelium because I do see white poking through in some spots. I was going to try carefully using peroxide again but not sure... any advice? Surprisingly, seems the pans Im trying are doing better that the cubes lol:-/
 
I also checked on the agar work and I think I finally got the plate, seems to have the right characteristics, ya?  :biggrin:
 

Attached Thumbnails

  • IMG_20200413_032932.jpg
  • IMG_20200413_033031.jpg
  • IMG_20200417_001024.jpg
  • IMG_20200417_001046.jpg
  • IMG_20200417_001115.jpg
  • IMG_20200417_031756.jpg
  • IMG_20200417_210700.jpg
  • IMG_20200420_203448.jpg
  • IMG_20200420_220802.jpg
  • IMG_20200420_220814.jpg


#16 scofos83

scofos83

    Mycophiliac

  • Free Member
  • 12 posts

Posted 23 April 2020 - 11:20 PM

 

So definitely getting what I asked for with learning and dialing new auto setups in lol :laugh: I had spawned to bulk sub of manure/straw, verm, and some gypsum. Cased with peat/verm/calcium carbonate. Misted and placed back in original closet. Due to drastic temp flux I made the decision (with girls approval haha) to move into the closet in the other room. After wiping down and cleaning I set it up the same which was 2 tubs stacked on each side with the humidifier between them, along with a heating pad both hooked to Inkbird controller. Everything seemed good once set up, however got case of noob paranoia coupled with the threads reminding over and over you cant just crank it up and walk away so I kept checking every hour or so and went to bed. I woke up in the am to inkbird alarm on humidity because excessive condensation had pooled in the tube leading to the control tub and blocked the humidity and air flow.  Im guessing lack of FAE, high humidity/temp, and/or a rushed setup introduced me to cobweb mold. And wow it IS impressively fast. The one tub went from small dime size to 3/4 of casing surface overnight and a second has small circular splotch.  I treated with Hydrogen peroxide spray and seemed to clear it up. I fixed the condensation issue by using a space heater in the actual room and tweaked the humidity system based on JOC tek idea with central reservoir.  I also got a Germ guardian air purifier that had $50 rebate:), UV, and Hepa to clean and add some flow. I first had it up on the top shelf on its side angled to blow downward towards the closet door and a small Vornado personal size fan to add some flow back up.
 
Ive since scrapped that too and changed setup which is 2 tubs on the right, 1 on left (ultimately woke to the tub totally covered in light gray cobweb so I tossed it), I added a new tub I got on the floor on top of my seedling mat to bump up ambient humidity, put the purifier on top still blowing a door (around 150cfm may be much), I put the heating pad on the shelf with a 4" AC Infinity duct fan on top with few inches of duct hose on the front and back. Essentially pushing air down past tubs to back of purifier, towards door, and back up again. Since making these changes I saw huge difference in temp and rh stability. Ive been able to hold around 75-80 deg @ 90-97% RH with what appears to be great evap rate, possibly too good? Its very humid in there but there is no vapor droplets on walls and sub surface "looks" dry, but not sure. Im nervous to keep humidity on surface high because I dont want that particular tub to succumb to the cobweb again. Today I see some growth on this PE tub in question, but hard to say if its cobweb or mycelium because I do see white poking through in some spots. I was going to try carefully using peroxide again but not sure... any advice? Surprisingly, seems the pans Im trying are doing better that the cubes lol:-/
 
I also checked on the agar work and I think I finally got the plate, seems to have the right characteristics, ya?  :biggrin:

 

I also had an issue when the tubes backed up and when I disconnected them some water must've ran back in down the sides. I believe the one is bacteria and the other other one either is or will be soon. Question is, can I cut that spot out and/or drain/soak up that excess water? I made adjustments so that cant happen again. Will update soon.

Attached Thumbnails

  • IMG_20200424_000927.jpg
  • IMG_20200424_000945.jpg





Like Mycotopia? Become a member today!