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Cleaning up dirty spores


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#21 peacefrog

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Posted 01 March 2017 - 08:38 PM

Update:

Cleaning up these spores is going as planned and moving in the right direction. Still some contamination, but getting much cleaner now. Below are the cleanest ones followed by a couple of the worst and dirtiest ones for your viewing pleasure lol.

TC plate:
IMG_0769.JPG

AM plate:
IMG_0770.JPG

HI plate:
IMG_0774.JPG

Contamination still strongly holding on to these:
IMG_0767.JPG IMG_0780.JPG
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#22 peacefrog

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Posted 07 March 2017 - 04:05 PM

Update:

 

After 3 transfers from the leading edge of the cleanest growth possible, I now have 3 clean plates, completely isolated from any/all competitors. It only took me a little over 2 weeks to get them cleaned up, and they were very dirty. If I had made a syringe with those dirty bastards and inoculated spawn or substrate directly, my results would have been terrible, even if I was a spore syringe master which I am not. This is why I love and use agar exclusively for all of my mushrooms.  

 

TC plate:

TC 3rd transfer.JPG

 

AM plate:

AM 3rd transfer.JPG

 

HI plate:

HI 3rd transfer.JPG

 

They are all now ready for LC, LI or grain inoculation. One could also continue isolating and testing each until a great culture is found. But I am only interested in a quick grow to replenish my spore bank, so these are going via LI into manure/BRF/verm cakes in PP5 containers. Final updates to come in the next few of weeks or so. 

 

 


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#23 Arathu

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Posted 07 March 2017 - 07:31 PM

Awesome......I have some work coming up too...................

 

A


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#24 peacefrog

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Posted 07 March 2017 - 07:44 PM

Thank you. I'm looking forward to seeing it!
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#25 whitethumb

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Posted 08 March 2017 - 11:48 PM

good to see you putting in a little work ;)
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#26 ChimX

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Posted 08 April 2017 - 11:13 AM

You guys and gals have convinced me entirely. Count me in!

1) 18 x 150/25mm Petri Dishes? Check✔
20170408_111937.jpg
20170408_111924.jpg
20170408_111911.jpg

2) Ginormous Acrylic Glove Box with 10 Liter PC Capacity? Check✔
20170116_173727.jpg
20170122_193250.jpg

3) Decent Collection of Dirty Spores? Check✔
20170408_114654.jpg

4) Slightest Notion of Agar Procedure?❌ Eek!...hmm...*cough cough*...(let's skip to 4B!)

4-B) Shameless Plan of Pestering Arathu, Catattack, MLB, Tails & Peacefrog to No End? Check✔

✅ OK, I'm all set!

Awesome thread Peacefrog. Thanks to everyone that's posted.

Metta
-ChimX

Edited by ChimX, 08 April 2017 - 11:24 AM.

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#27 dead_diver

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Posted 08 April 2017 - 01:01 PM

Has anyone tried pouring agar over the myc and contam in a dish to see if myc would grow up through the agar but leave the contam behind?
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#28 coorsmikey

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Posted 08 April 2017 - 01:13 PM

Has anyone tried pouring agar over the myc and contam in a dish to see if myc would grow up through the agar but leave the contam behind?

I have tried that and the hot agar killed everything on the plate for the most part. It did end with contamination later. I don't know if it was bacteria that survived the heat or was introduced later.
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#29 peacefrog

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Posted 08 April 2017 - 02:42 PM

ChimX,
Thank you for the nice words. And ask away, that's what this thread is all about. Learning and sharing.

Mikey,
Did you try the sandwich tek several times and every time it failed? I have never attempted it and only have read about it. I am just stuck in my ways of cleaning up the old fashioned way lol. I would love it if you explained your experience on this thread, as it might possibly help others if that tek is crap.

If it is truly a crappy tek, perhaps the op can be edited and remove my comments on it? I don't want to pass along bad info. I just assumed it was a valid procedure from all of my reading. And you know what they say, "when you assume you make an a@% of yourself lol.

Edited by peacefrog, 08 April 2017 - 03:02 PM.


#30 Arathu

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Posted 08 April 2017 - 04:36 PM

I'll have a few days off from an insane work schedule and will probably be getting bitched at by the family for spending too much time on my fungal friends......none the less a blue agar pour is imminent....

 


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#31 Juthro

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Posted 08 April 2017 - 05:01 PM

Really nice thread, PeaceFrog. Nice pics, and well written :)
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#32 coorsmikey

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Posted 08 April 2017 - 06:55 PM

ChimX,
Thank you for the nice words. And ask away, that's what this thread is all about. Learning and sharing.
Mikey,
Did you try the sandwich tek several times and every time it failed? I have never attempted it and only have read about it. I am just stuck in my ways of cleaning up the old fashioned way lol. I would love it if you explained your experience on this thread, as it might possibly help others if that tek is crap.
If it is truly a crappy tek, perhaps the op can be edited and remove my comments on it? I don't want to pass along bad info. I just assumed it was a valid procedure from all of my reading. And you know what they say, "when you assume you make an a@% of yourself lol.

The same subject was brought up years ago and I only tried pouring agar over a contaminated dishes to politely point out that the thermal death limit of mycelium speaking in temperature is much lower than thermal death of many bacteria. I suppose you could try and let the agar cool near 100 f degrees and see if will still pour for you. I have never measured the temp of agar when I go to pour but I know I must use something to keep from burning my hands. Food is kept at 145f to keep when being served and I can generally eat that right away with burning my mouth so I have just "assumed" that when pouring agar it is much hotter than 145f. When I attempted this years ago it was only to prove that point. I don't think anything need to be edited and the idea could be further explored once the lowest possible temp to pour agar is established.

Edited by coorsmikey, 08 April 2017 - 08:39 PM.

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#33 peacefrog

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Posted 10 April 2017 - 06:53 AM

Thank you, Mikey for adding your experience to this thread.

I doubt I will ever try it, as my old ways seem to work very well for me personally. But for others out there, it might be a viable avenue to take while cleaning up bacterial agar. I hope others chime in with their experience with it.

Thank you, Juthro for the kind words!

Edited by peacefrog, 10 April 2017 - 06:55 AM.

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#34 sandman

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Posted 10 April 2017 - 07:27 AM

You can do a sandwich of agar without pouring hot agar on the plate. You simply cut and lift a square of agar from another plate. Never really messed with it personally because haven't really had the need.


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#35 CatsAndBats

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Posted 10 April 2017 - 07:42 PM

@peacefrog, what's your per jar recipe for agar again? Sometimes I just need a couple of jars made up not a whole batch. Great thread! You're the best.


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#36 peacefrog

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Posted 14 April 2017 - 05:13 AM

You da man with that new name lol.

I usually make up enough to 6-8 jars and use a heaping teaspoon per jar filled with 15 ml. If I only make a couple of jars, I store the left over mix in a sealed ziplock bag. This doesn't work so well with MEA, but all the other recipes I use, it does well.

I have tried in the past to measure out just enough for 1 jar with mixed results. My scale is a cheap one from Office Max and doesn't even register until 2 grams. But if I make up a batch of PDA using 5 grams instant potatoes, 5 grams of agar agar, 2 grams of dextrose, the results are much better from my experience. So I just do that and store for later.

Than you Sandman for chiming in as well.

Edited by peacefrog, 14 April 2017 - 05:19 AM.

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#37 peacefrog

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Posted 02 May 2017 - 10:22 AM

Final update and all grown up:

 

TC:

28. TC Pins.JPG 29. TC Pins.JPG 30. TC Pins.JPG 37. TC Mature.JPG 38. TC Mature.JPG 39. TC Mature.JPG

 

HI:

31. HI Pins.JPG 32. HI Pins.JPG 33. HI Pins.JPG 40. HI Mature.JPG 41. HI Mature.JPG 42. HI Mature.JPG

 

AM:

34. AM Pins.JPG 35. AM Pins.JPG 36. AM Pins.JPG 43. AM Mature.JPG 44. AM Mature.JPG 45. AM Mature.JPG

 

My spore collection has been renewed.  

 


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#38 Arathu

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Posted 02 May 2017 - 10:27 AM

Schweet!

 

I think I'm going to have to learn to do short threads.................Nice work PF..........

 

I'm headed into edibles big time myself............so new days and new adventures to come.........

 

Man I love mushrooms............

 

A


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#39 peacefrog

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Posted 02 May 2017 - 06:04 PM

Thank you for the kind words, Arathu.

Edibiles are awesome and my wife and I love eating some fresh oysters sautéed with butter and a bunch of veggies!

I also share your love for our fungal friends.
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#40 MLBjammer

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Posted 02 May 2017 - 06:32 PM

Schweet!

 

I think I'm going to have to learn to do short threads.................Nice work PF..........

 

I'm headed into edibles big time myself............so new days and new adventures to come.........

 

Man I love mushrooms............

 

A

You, like me, are not allowed to do short threads, man.  It is just impossible for us, lol.

 

Beautiful little trays, PF.  Finesse growing at its finest.


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