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DrMycHunter
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First time with Agar and everything else
#26791633 - 06/29/20 08:43 AM (3 years, 7 months ago) |
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Hello all. This is my first try at cultivation. After a bunch of reading I decided to jump right in to agar from the start. I went with a pre made MEA I got off ebay (35.7% Malt Extract, 1.8% Peptone, 35.7% Dextrose, 26.8% Agar Agar) and added 10g of that mix to 400 ml of water and a little dab of food coloring. I don't own a microwave so I was going to heat the water to mix the solution but I spaced it and forgot that step. I PC'd it at 16 psi for 25 mins and let it cool to ~47C before I poured.
After my plates were poured I followed BOD's agar tek write-up and swiped 20 plates using 4 different syringes.
Here's what I've ended up with so far....my SAB use needs some serious improvement haha
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InnerWisdom



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Re: First time with Agar and everything else [Re: DrMycHunter]
#26791658 - 06/29/20 09:01 AM (3 years, 7 months ago) |
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First one looks like mycelium but why are there so many different spots?
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Kmacmo
The aborted pin



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Re: First time with Agar and everything else [Re: DrMycHunter]
#26791660 - 06/29/20 09:04 AM (3 years, 7 months ago) |
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Good job you have some mycelium on those plates  The agar/nutes mix hasn't dissolved properly wich isn't really an issue, but next time disovle properly and makes it easyer to see what's going on.
That big blue spot is penicillin Mold. Learning agar is the best thing you can ever do :P
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DrMycHunter
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Re: First time with Agar and everything else [Re: InnerWisdom]
#26791680 - 06/29/20 09:18 AM (3 years, 7 months ago) |
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I have poor fine hand control, I was assuming I just spooged too much liquid from the syringe.....I also didn't shake up my syringes before use so maybe that as well? Like I said I'm just starting so I'm pretty clueless hahahaha. Thanks all for the quick replies.
Just trash the plates with Pen growing I'm assuming?
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DrMycHunter
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Re: First time with Agar and everything else [Re: DrMycHunter]
#26791682 - 06/29/20 09:20 AM (3 years, 7 months ago) |
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My Golden Teacher plates are a complete mess, I'll get some pics up for shits and giggles. Btw Thanks for all those videos Bod they were super helpful, I had a fun time building my SAB and putting it to use.
Edited by DrMycHunter (06/29/20 10:44 AM)
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Roger Clemency
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Re: First time with Agar and everything else [Re: DrMycHunter]
#26791823 - 06/29/20 10:43 AM (3 years, 7 months ago) |
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Germination plates will often have other stuff on them which is fine, you just cut out a little wedge with clean mycelium and transfer that.
If you’re noticing satellite colonies of mold or bacteria, meaning bad stuff is showing up in an area where you didn’t swipe spores or transfer something from another plate, then those are getting in when you’re opening your dish, or they’re getting in from not having plates wrapped etc. and then you need to take a look at your SAB technique.
-------------------- Sour grapes, sweet revenge Heaven starts right where hell ends
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DrMycHunter
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Re: First time with Agar and everything else [Re: Roger Clemency]
#26791883 - 06/29/20 11:16 AM (3 years, 7 months ago) |
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Quote:
Roger Clemency said: Germination plates will often have other stuff on them which is fine, you just cut out a little wedge with clean mycelium and transfer that.
By that do you mean I could actually clean up those plates with Pen growth?
Quote:
Roger Clemency said: If you’re noticing satellite colonies of mold or bacteria, meaning bad stuff is showing up in an area where you didn’t swipe spores or transfer something from another plate, then those are getting in when you’re opening your dish, or they’re getting in from not having plates wrapped etc. and then you need to take a look at your SAB technique.
Thanks for the reply and the info, very appreciated. It was definitely my SAB technique I'm sure. I forgot to spray down my box on the first 5 plates which were the GTs and those are a mess, the others I'm pretty sure my hand crossed over the plates while working. I was a little nervous but now that I've done it once I should be fine going forward.
I think I wrapped them pretty well, I used ClingWrap and did three full rotations (some a little more if it snapped mid wrap)
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Roger Clemency
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Re: First time with Agar and everything else [Re: DrMycHunter]
#26791902 - 06/29/20 11:33 AM (3 years, 7 months ago) |
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Yeah you’d just cut a little piece out with the myc you want and drop that onto a new clean plate to grow out. You don’t want to try to cut out the contaminant if that’s what you meant though. It could work but could leave spores or little filaments that reached out farther than it looked.
Some people do actually use their SAB totally dry. I just spray mine with a light mist of water and wait a couple minutes before starting. Once you do it a couple times and are more comfortable it should get better. Watch some videos of you can of people doing SAB work.
-------------------- Sour grapes, sweet revenge Heaven starts right where hell ends
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DrMycHunter
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Re: First time with Agar and everything else [Re: Roger Clemency]
#26792083 - 06/29/20 01:21 PM (3 years, 7 months ago) |
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Quote:
Roger Clemency said: Yeah you’d just cut a little piece out with the myc you want and drop that onto a new clean plate to grow out. You don’t want to try to cut out the contaminant if that’s what you meant though. It could work but could leave spores or little filaments that reached out farther than it looked.
I did mean off the Myc, thanks for the clarification. I'm looking forward to this fun.
Quote:
Roger Clemency said: Some people do actually use their SAB totally dry. I just spray mine with a light mist of water and wait a couple minutes before starting. Once you do it a couple times and are more comfortable it should get better. Watch some videos of you can of people doing SAB work.
Hahaha I think I've been watching these videos more than some watch porn....
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DrMycHunter
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Re: First time with Agar and everything else [Re: DrMycHunter]
#26799442 - 07/02/20 01:58 PM (3 years, 7 months ago) |
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Hahaha No idea what I did to the GT plates, they were the first 5 I made and the only ones to have this issue, however. Did my first transfers today on PESA and Malabar, hopefully they look alright and I didn't mess up too bad in practice. Time will tell I guess 
Edited by DrMycHunter (07/02/20 02:00 PM)
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Roger Clemency
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Re: First time with Agar and everything else [Re: DrMycHunter]
#26799666 - 07/02/20 03:25 PM (3 years, 7 months ago) |
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Yooo you landed white side up on that first blue plate lol it looks good though from the bit I can see. Can't tell whats up with the Gt though.
-------------------- Sour grapes, sweet revenge Heaven starts right where hell ends
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DrMycHunter
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Re: First time with Agar and everything else [Re: Roger Clemency]
#26799744 - 07/02/20 04:08 PM (3 years, 7 months ago) |
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Quote:
Roger Clemency said: Yooo you landed white side up on that first blue plate lol it looks good though from the bit I can see. Can't tell whats up with the Gt though.
Hahaha I tried so hard to get em all myc side down....a couple just refused so I went with it.
Round two of pouring plates went way better, I let the agar solution cool off even more this time which seemed to work really well, I also remembered to heat it prior to PC'n to mix it. I tossed all those GT plates today....way beyond me at this point lol.
Got some spores prints to try working with at some point since I'm having such a good time with all this
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Roger Clemency
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Re: First time with Agar and everything else [Re: DrMycHunter]
#26800030 - 07/02/20 06:28 PM (3 years, 7 months ago) |
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Nice. I hate when they fall white side up lol. Sometimes i'll nudge them over, sometimes just leave them. They grow out real quick anyway
-------------------- Sour grapes, sweet revenge Heaven starts right where hell ends
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DrMycHunter
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Re: First time with Agar and everything else [Re: Roger Clemency]
#26800956 - 07/03/20 07:50 AM (3 years, 7 months ago) |
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Quote:
Roger Clemency said: Nice. I hate when they fall white side up lol. Sometimes i'll nudge them over, sometimes just leave them. They grow out real quick anyway
You weren't joking about quick, the Myc already covered the transfer pieces I cut out! So exciting! Mawhahaha


The second pic is the MC from my first post, I ran out of plates for transfers but thankfully have more coming in today....need to order a case next time lol
Edited by DrMycHunter (07/03/20 10:36 AM)
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DrMycHunter
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Re: First time with Agar and everything else [Re: DrMycHunter]
#26809348 - 07/07/20 03:42 PM (3 years, 7 months ago) |
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Well dang, my order of petri dishes got pushed back until Sat and I need to do some transfers....went ahead and ordered a case of 500 so this never happens again lol
Should I pack them up and put them in cold storage until Sat?
Edited by DrMycHunter (07/07/20 03:43 PM)
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DrMycHunter
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Re: First time with Agar and everything else [Re: DrMycHunter]
#26817201 - 07/11/20 12:15 PM (3 years, 7 months ago) |
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Alright got my case of dishes, now it's time to weed through the messes, just about what I was expecting after I almost pulled over my stack of transfers right as I was just about done before I taped them up....sigh. Any suggestions would be awesome, I went through the contam threads to try and Id stuff but I really don't trust my eyes with all this yet.
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DrMycHunter
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Re: First time with Agar and everything else [Re: DrMycHunter]
#26817231 - 07/11/20 12:26 PM (3 years, 7 months ago) |
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I had trouble getting the Golden Teacher syringe I had to do anything on Agar so I just decided to inoculate some oat spawn jars (followed Bod's Oat prep tek) to try getting a few grains I could use.
I can't tell if things are progressing nicely or not, some of it looks to be contaminated but like I said I don't trust my eyes yet.
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InnerWisdom



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Re: First time with Agar and everything else [Re: DrMycHunter]
#26817337 - 07/11/20 01:10 PM (3 years, 7 months ago) |
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Looks totally fine. I don't know what exactly you are trying to highlight there, but it all looks normal and nicely uniform. Mycelium has many different looks when it is colonizing depending on the genetics and environment I think.
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DrMycHunter
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Re: First time with Agar and everything else [Re: InnerWisdom]
#26817365 - 07/11/20 01:21 PM (3 years, 7 months ago) |
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I was just tracking growth with the marker, thanks for the reply appreciated
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DrMycHunter
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Re: First time with Agar and everything else [Re: DrMycHunter]
#26820893 - 07/13/20 11:17 AM (3 years, 7 months ago) |
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Shook a few of the jars last night, I was surprised to see them recovered already today.
Edited by DrMycHunter (07/13/20 11:18 AM)
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