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MycPsilo
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First Agar Experiment
#26512285 - 03/02/20 04:37 AM (3 years, 10 months ago) |
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If this is in the wrong place please let me know. SO These are the first plates I have ever poured or inoculated. Used a spore syringe. 1-2 drops. Is everything looking ok? Too early to tell..? How large do you let it get before starting any transfers?
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Sockadin



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Re: First Agar Experiment [Re: MycPsilo]
#26512307 - 03/02/20 05:06 AM (3 years, 10 months ago) |
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Large enough to see what clean growth is happening in the dish. Between a quarter and a silver dollar.
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Ombisha
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Re: First Agar Experiment [Re: Sockadin]
#26512324 - 03/02/20 05:39 AM (3 years, 10 months ago) |
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Hard to tell from those pictures. I see two spots in the first picture - are those bubbles or something growing?
Quote:
Sockadin said: Large enough to see what clean growth is happening
yeah, since you used spore solution, there will probably be contamination present. For the first 1-2 transfers, you will just want to transfer the mycelium that will be farthest from the contamination. After you get rid of the contamination, you will want to isolate the strain. That means that you will want to grow it out to about 1/2 - 3/4 of the dish, then take a small piece from the leading edge with the nicest growth.
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MycPsilo
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Re: First Agar Experiment [Re: Ombisha]
#26512342 - 03/02/20 06:13 AM (3 years, 10 months ago) |
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Quote:
Sockadin said: Large enough to see what clean growth is happening in the dish. Between a quarter and a silver dollar.
Quote:
Ombisha said: Hard to tell from those pictures. I see two spots in the first picture - are those bubbles or something growing?
Quote:
Sockadin said: Large enough to see what clean growth is happening
yeah, since you used spore solution, there will probably be contamination present. For the first 1-2 transfers, you will just want to transfer the mycelium that will be farthest from the contamination. After you get rid of the contamination, you will want to isolate the strain. That means that you will want to grow it out to about 1/2 - 3/4 of the dish, then take a small piece from the leading edge with the nicest growth.
Yes the second spot in the first pic is just bubbles. I guess my biggest worry is being able to even spot or identify any possible contamination.
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MycPsilo
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Re: First Agar Experiment [Re: Sockadin]
#26512345 - 03/02/20 06:17 AM (3 years, 10 months ago) |
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Quote:
Sockadin said: Large enough to see what clean growth is happening in the dish. Between a quarter and a silver dollar.
Ok...so at this size, identifying possible contamination will be to small to tell?
Also, the growth that is going on in there seems kinda "fuzzy"...Is this because I am using a nutrient rich agar premix? Because its from MS?
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shevanel
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Re: First Agar Experiment [Re: MycPsilo]
#26512395 - 03/02/20 07:35 AM (3 years, 10 months ago) |
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just here to pick up info
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MycPsilo
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Re: First Agar Experiment [Re: shevanel]
#26513104 - 03/02/20 03:44 PM (3 years, 10 months ago) |
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Edited by MycPsilo (03/03/20 10:32 AM)
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shevanel
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Re: First Agar Experiment [Re: MycPsilo]
#26513120 - 03/02/20 03:54 PM (3 years, 10 months ago) |
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i cant get a sharpie to write on a petri. what kind of marker do you use?
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Sockadin



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Re: First Agar Experiment [Re: shevanel]
#26513130 - 03/02/20 04:03 PM (3 years, 10 months ago) |
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Black medium point
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MycPsilo
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Re: First Agar Experiment [Re: shevanel]
#26513131 - 03/02/20 04:04 PM (3 years, 10 months ago) |
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Just a regular sharpie med point. Maybe its your plates?
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shevanel
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Re: First Agar Experiment [Re: MycPsilo]
#26513135 - 03/02/20 04:07 PM (3 years, 10 months ago) |
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yea. the ezbio research ones from amazon
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rm1024
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Re: First Agar Experiment [Re: shevanel]
#26513138 - 03/02/20 04:10 PM (3 years, 10 months ago) |
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looks good from my house.
@schevanel: i use regular sharpie brand. have you tried a fresh one? if your dishes have alcohol on them it can smear it. works for me on poly and glass plates.
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Ignorantape
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Re: First Agar Experiment [Re: rm1024] 1
#26513341 - 03/02/20 06:05 PM (3 years, 10 months ago) |
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Your plates are looking good there. No obvious signs of contam. Just be patient and leave it for a few days.
And you'll get that fuzzy growth on early plates. It's called tomentose growth as opposed to the rhizomorphic growth that you'll see when you have cleaner transfers. I believe it's different because at this early stage you've got all the different strains of myc growing over each other.
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MycPsilo
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Re: First Agar Experiment [Re: MycPsilo]
#26514382 - 03/03/20 10:33 AM (3 years, 10 months ago) |
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Eclipse3130
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Re: First Agar Experiment [Re: MycPsilo]
#26514394 - 03/03/20 10:40 AM (3 years, 10 months ago) |
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If you are using a certain pre mix agar from Amazon you almost always will get extra fuzzy growth, it makes it hard to see rhizomorph underneath it. Comes from the excess sugar/nutrient, and some strains are naturally tomentose, it's very hard to tell with a heavy nutrient rich agar, but it still grows just fine
-------------------- "In The Material World One seeks retirement and grows Old In The Magical World One seeks Enlightenment and grows Wiser In The Miraculous World One seeks nothing and grows Lighter As we all tread the Homeward Path we will explore many Realms And one day... we will all Realize that all experiences are Simply Different ways in which The All-That Is Perceives Itself"
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MycPsilo
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Quote:
Eclipse3130 said: If you are using a certain pre mix agar from Amazon you almost always will get extra fuzzy growth, it makes it hard to see rhizomorph underneath it. Comes from the excess sugar/nutrient, and some strains are naturally tomentose, it's very hard to tell with a heavy nutrient rich agar, but it still grows just fine
Yeah that was what I thought.
I am using a RAK MEA premix from a supply store.
I had heard that it was pretty nutrient rich. I used one gram less of the premix than what was recommended.
I was wondering how much I could play with this. If I use too much less that would make my plates softer right?
This being my first time I wanted to just use the mix. I think once that is used up I will switch to just making my own.
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MycPsilo
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Re: First Agar Experiment [Re: MycPsilo]
#26514400 - 03/03/20 10:45 AM (3 years, 10 months ago) |
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Also...will that tomentose growth go away as I make transfers? Become more rhizo? Or will that just be the nature of the beast with the mix that I am using?
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Eclipse3130
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Re: First Agar Experiment [Re: MycPsilo]
#26514412 - 03/03/20 10:52 AM (3 years, 10 months ago) |
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Yeah to fix it you basically have to make your own eventually with LME and agar, the pre mix brands usually have unnecessary dextrose added which probably is the cause to the fuzzy growth.
It's fine though, reducing it won't do much help besides making your agar less firm.
Transfering tomentose will most likely produce more tomentose, in this case you can't really say for sure the true nature of the growth with excess nutrients but I would transfer some anyway
-------------------- "In The Material World One seeks retirement and grows Old In The Magical World One seeks Enlightenment and grows Wiser In The Miraculous World One seeks nothing and grows Lighter As we all tread the Homeward Path we will explore many Realms And one day... we will all Realize that all experiences are Simply Different ways in which The All-That Is Perceives Itself"
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MycPsilo
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Ok...haha...a bit discouraging but it will be ok.
I was hoping the tomentose growth might have to do with multiple strains and a nutrient agar but would remedy itself through isolation.
Thank you for pointing out it will probably still be fuzzy...headed to Amazon to buy what I need to mix my own agar.
Im sure there is a good recipe here somewhere.
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shevanel
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Re: First Agar Experiment [Re: Eclipse3130] 1
#26514427 - 03/03/20 11:01 AM (3 years, 10 months ago) |
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i too am using rak media mea and also have tomentose growth on some but not all plates.
this is actually a t3 not t2

then most of the early plates were nearly half n half
Edited by shevanel (03/03/20 11:02 AM)
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MycPsilo
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Re: First Agar Experiment [Re: MycPsilo]
#26514432 - 03/03/20 11:04 AM (3 years, 10 months ago) |
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In the meantime...while I play around with isolating...What would be my best method of getting some grain jars going?
Would it be alright to take a clean wedge of my next transfer and drop it into a grain jar? Then G2G?
I know it wont be as good, I am just wanting to get a tub going. Possibly clone the best out of that flush and have that start my next experiments.
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InfiniteDreams


Registered: 10/25/19
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Re: First Agar Experiment [Re: Eclipse3130] 1
#26514435 - 03/03/20 11:04 AM (3 years, 10 months ago) |
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To a degree, I have found that repetition of transfering tomentose, especially towards a lower nutrient agar, can allow the tomentose growth to switch towards rhizo.
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