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Offlinemoshe1

Registered: 08/29/15
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Help me understand my agar
    #27325787 - 05/27/21 10:01 PM (18 days, 5 hours ago)

Here are some of my agar dishes currently growing out. This is my first ever batches of agar, as in i've done germ plates, transfers 1 + 2s.  i'm yet to have put any agar to grain yet.

I'll label them respectively and leave it open ended- because i'm trying to understand whats going on and make decisions based on some knowledge and im having a hard time. My confusion is coming from having sporadic growth, the myc is all over the place and makes it hard for me to understand what im looking at. Theres aerial growths, thin and almost transparent growths, fuzzy areas, flat areas. Spots that seem to be growing out in an even fashion, and some that look like a jungle. Also note, I've had no discolorations, even my germ plates from a month ago are still white although unusual looking.


trying not to ask too many questions with new threads but my search isn't yielding any help that im looking for- rather confusing me more with all the mixed answers i am reading on every thread.
https://www.shroomery.org/forums/showflat.php/Number/26988516#26988516 ;  this thread/response from RR is the only thing that really looks similar to mine that i could possibly take advice for my situation without uncertainties

Using bods LME for my agar.  About 400 ML water, 10 grams AGAR to roughly 7.5 grams LME for every batch of 20 plates. 

First sign of rhizo growth anywhere on any plate. Showed up on a T1 plate way later, regrowing out of an area in the center.


Mazy germ, t1 and t2. A large portion of my plates grew out thin and translucent like this before getting any kind of thickness



A couple LGT Germs and their T1s, took over the whole plate already. Had too many plates to transfer so i let these grow out, got handful more that look like them.


BBM germ, t1 and t2


BBM T1 plates that are almost grown out completely


BBM Germ t1 and t2


Close up of a T1 plate fully grown out. Still no signs of "strong rhizo growth" anywhere to be found- anywhere



most organized / thick myc from the bunch across all varieties, but still very fuzzy


Edited by moshe1 (05/28/21 01:46 AM)


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Offlineverum subsequentis
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Re: Help me understand my agar [Re: moshe1]
    #27325792 - 05/27/21 10:09 PM (18 days, 5 hours ago)

Spore syringe?


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InvisibleFeasoghorm

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Re: Help me understand my agar [Re: moshe1]
    #27325799 - 05/27/21 10:13 PM (18 days, 5 hours ago)

Yeah pretty much all them plates cept for the germ plates look odd. Well, germ plates look odd in general, but i thnk that you have a contaminant fungus/mold blended in with your myc. I used to say "seamless", but it aint really seamless if you can tell it's fucked haha.


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Offlinemoshe1

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Re: Help me understand my agar [Re: verum subsequentis]
    #27325804 - 05/27/21 10:15 PM (18 days, 5 hours ago)

Quote:

verum subsequentis said:
Spore syringe?




all of these are off print/swab

i do have a couple from spore solution that are a little behind- but more-or-less still hard to read like these are.




Edited by moshe1 (05/27/21 10:22 PM)


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Offlineverum subsequentis
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Re: Help me understand my agar [Re: moshe1] * 1
    #27325807 - 05/27/21 10:18 PM (18 days, 5 hours ago)

Middle plate in second pic.
Bottom two in last pic.

These are your best chances at getting a clean culture.


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InvisibleFeasoghorm

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Re: Help me understand my agar [Re: verum subsequentis]
    #27325812 - 05/27/21 10:21 PM (18 days, 5 hours ago)

I've also seen odd plates like this from improperly nuted grain soak agar. However i dnt thnk there is anything wrong with your mix.

Looks like he transferred from middle plate 2nd pic to 3rd plate 2nd pic.
Bottom right in last pic doesnt look too bad.


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Offlinemoshe1

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Re: Help me understand my agar [Re: verum subsequentis]
    #27325814 - 05/27/21 10:23 PM (18 days, 5 hours ago)

Quote:

Feasoghorm said:
I've also seen odd plates like this from improperly nuted grain soak agar. However i dnt thnk there is anything wrong with your mix.

Looks like he transferred from middle plate 2nd pic to 3rd plate 2nd pic.
Bottom right in last pic doesnt look too bad.





what are the odds of this happening on every batch? I mean, im only like 4 batches of agar in sure- but whats riding on every single plate i germ or transfer?  I let my flowhood in the clean-room run with the door shut for like, 3 hours, sometimes up to 5 to scrub the air before i work. Mind you this room is only for mixing/spawning my tubs and doing anything that requires the flowhood- and i do my best to keep sterile technique when i work.  of course im still a noob with agar- but i still need to identify problems to fix em.

if all is fucked in these pics than im pretty concerned- cause that would indicate that my whole shelf of agar plates are contaminated with mold?


Quote:

verum subsequentis said:
Middle plate in second pic.
Bottom two in last pic.

These are your best chances at getting a clean culture.




noted.


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Edited by moshe1 (05/27/21 10:27 PM)


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InvisibleFeasoghorm

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Re: Help me understand my agar [Re: moshe1]
    #27325829 - 05/27/21 10:36 PM (18 days, 5 hours ago)

Well im not for certain wats up with your plates, although i wish i cud be. It's jst my best guess.

I've never had a contaminated syringe so i dnt knw what to say bout that, but ive run into many a dirty print/swab. I expect it.
It can be difficult to differentiate freahly germinated cube spores and freshly germinated aspergilus.
Do you scrape spores off of prints with the print above the plate? This is a contam laden practice.

There cud be something happening pre-inoculation. Do you set aside a control plate from each batch? Do you wait 2wks b4 using fresh plates? If i can i will, cuz ive had shit show up 2wks from pour.

When you work in your SAB, do you wear a mask? Are you right up on it looking through the top to work? Or in front of it?

Also, are you using the same print/swab for all these? Like, are these genetics basically all from the same shroom?

Do you wear gloves?


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Offlinemoshe1

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Re: Help me understand my agar [Re: Feasoghorm]
    #27325922 - 05/28/21 12:11 AM (18 days, 3 hours ago)

Quote:

Feasoghorm said:
Well im not for certain wats up with your plates, although i wish i cud be. It's jst my best guess.

I've never had a contaminated syringe so i dnt knw what to say bout that, but ive run into many a dirty print/swab. I expect it.
It can be difficult to differentiate freahly germinated cube spores and freshly germinated aspergilus.
Do you scrape spores off of prints with the print above the plate? This is a contam laden practice.

There cud be something happening pre-inoculation. Do you set aside a control plate from each batch? Do you wait 2wks b4 using fresh plates? If i can i will, cuz ive had shit show up 2wks from pour.

When you work in your SAB, do you wear a mask? Are you right up on it looking through the top to work? Or in front of it?

Also, are you using the same print/swab for all these? Like, are these genetics basically all from the same shroom?

Do you wear gloves?




My process:

After rooms air has been scrubbing for a few hours at least, i go in. remove shirt i was just wearing outside/during the day, spray arms up to shoulders up with ISO, gloves on, mask on. Then with my alchohol towel i wipe down all surfaces, and mist ISO on the filter too.  Then ring unopened sleeve of plates over, tools, and whatever else.

then i go pick out what im going to transfer or germ, bring those as well.  As for technique, always wiping with ISO before bringing in flow, always flame sterilizing tools when needed. I always focus on not letting my hands get in-between any flow whatever it is im working on.


I've been prepping my agar and pouring it the day of, or next day- and usually inoculate the next day after pouring, unless i've been at the office long enough for it to cool, but thats only happened once so far.

i have not set aside any control plates on purpose- but i have had extra plates that have been poured i didnt use because of condensation- and those have stayed clean even without being wrapped with parafilm and moved around carelessly. They look spotless, and thats with almost zero care, they're just sitting in the lab rn still clean-  my plates i've worked with are exponentially handled with more carefulness.

i've never waited longer than a day or two after pouring my plates-  after i pour i just put the sleeve back over the stack without sealing them until i work with them again the next day or two. I have a bottle of agar in the fridge now waiting to be poured, maybe ill pour early and give it more time. At this point i'm not sure what to do.

also, they are all different! thats why- its so confusing when they all show oddness.  I got Blue meanies from print/swab, Mazy from print/swab, LGT from print/swab which are all pictured above.
They are all similar in "oddness" lol...

I really need another friend in this hobby to come and look at my stuff in person and be able to conversate with lol


Edited by moshe1 (05/28/21 12:31 AM)


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InvisibleFeasoghorm

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Re: Help me understand my agar [Re: moshe1]
    #27326013 - 05/28/21 12:46 AM (18 days, 3 hours ago)

Shit dude...idk


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Offlinemoshe1

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Re: Help me understand my agar [Re: Feasoghorm]
    #27326053 - 05/28/21 01:30 AM (18 days, 2 hours ago)

all good man- when i figure out whats happening i'll definitely post it. Like i said- no one has given me definitive answers yet and searching on the forums will yield you in so much mixed feedback you will catch yourself going back and forth on what you think.

when i get a chance to test a couple plates the answer will be clear on the contam side of things.

bump for more eyes - here are some close ups of germ plates and transfer ones as clear as i could get them.  you could really zoom in on em if you open in a new tab and see.

https://www.shroomery.org/forums/showflat.php/Number/26988516#26988516 this response from RR to someone with the closest lookign things to my plates is all thats giving me hope lmao



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Edited by moshe1 (05/28/21 01:42 AM)


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Offlineverum subsequentis
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Re: Help me understand my agar [Re: moshe1]
    #27326324 - 05/28/21 09:01 AM (17 days, 18 hours ago)

Yowzers. You've got some wild shit going on. I'd start some new spores.  Use a loop and streak a few plates. Update here with pics as you go and we can help you understand where things are going wrong.


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OfflineBenson
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Re: Help me understand my agar [Re: verum subsequentis]
    #27326368 - 05/28/21 09:47 AM (17 days, 18 hours ago)

Not sure how helpful this is because some of your plates have thick growth of something but as for the wispy ones I encountered that a few months ago. I upped my LME and added 1g yeast and the growth got thicker/healthier. I would definitely suggest new spores in any case, cause that shit looks funky...

Wispy


Blue is more LME and +1g yeast


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OfflinemushboyM
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Re: Help me understand my agar [Re: Benson]
    #27326369 - 05/28/21 09:49 AM (17 days, 18 hours ago)

i 3rd for new spores.

or do a proper streak with the ones ya got.. but if you want quality myc relatively painlessly id just get new spores.


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Offlineverum subsequentis
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Re: Help me understand my agar [Re: mushboy]
    #27326372 - 05/28/21 09:50 AM (17 days, 18 hours ago)

Fourthed


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OfflineFeedYourMind
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Re: Help me understand my agar [Re: Benson]
    #27326514 - 05/28/21 11:56 AM (17 days, 15 hours ago)

Quote:

Benson said:
Not sure how helpful this is because some of your plates have thick growth of something but as for the wispy ones I encountered that a few months ago. I upped my LME and added 1g yeast and the growth got thicker/healthier. I would definitely suggest new spores in any case, cause that shit looks funky...

Wispy


Blue is more LME and +1g yeast





I had the same experience as you and OP. I had thin and wispy growth on agar and upped my nutes to the standard 10g of MEA per 500ml/water. I also add in 1-2g of nutritional yeast and that made ALL the difference in the world for me. Also, I started using .5g-1g of soy peptide per 500ml water and that has also improved the growth speed - especially when starting from spores.


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InvisibleFeasoghorm

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Re: Help me understand my agar [Re: FeedYourMind]
    #27326617 - 05/28/21 01:49 PM (17 days, 14 hours ago)

Interesting :strokebeard:
Getting many reports of this from either too much or not enough nutes in the mix. 10A/10LME/500ml is the standard. Didnt thnk a 2.5 gram reduction in LME and 100ml reduction in water to the mix wud fux wit it that much.


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Offlinemoshe1

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Re: Help me understand my agar [Re: Feasoghorm]
    #27326844 - 05/28/21 04:58 PM (17 days, 10 hours ago)

i have some fresh melmaks that just germinated waiting for transfers. i'm gonna transfer a few of these wild looking ones and see what happens anyway and update w/ pics.

i've been on the same thought dude, some say lower nutes some say up them and they are both to remedy the same issues lmao. gotta love it.

im a little disappointed but i guess it is what it is, all my plates have that kind of vertically growing fuzz some more some less. wonder what it could be, across all the different varieties i have too. my rooms air is scrubbing as we speak to pour more agar.

interesting thing is that this stuff isnt changing color or shape or anything, just bright white like the myc. all the germ plates look like they have white cotton on them


thanks for all the feedback. any suggestions or favorites for what variety i should put to agar to catch me up?


--------------------


Edited by moshe1 (05/28/21 05:08 PM)


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OfflineBenson
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Re: Help me understand my agar [Re: Feasoghorm]
    #27327076 - 05/28/21 08:44 PM (17 days, 7 hours ago)

Quote:

Feasoghorm said:
Interesting :strokebeard:
Getting many reports of this from either too much or not enough nutes in the mix. 10A/10LME/500ml is the standard. Didnt thnk a 2.5 gram reduction in LME and 100ml reduction in water to the mix wud fux wit it that much.




I'm convinced at this point it's the nutrient level of the LME... I have nothing to base that off other than process of elimination tho. Distilled water, +10g agar (one of the two commonly used brands), +10g LME works great for some like D3, but was trash for me across multiple strains and swabs. Instant results when I upped the neuts :shrug:


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Re: Help me understand my agar [Re: Benson]
    #27327118 - 05/28/21 09:10 PM (17 days, 6 hours ago)

Hey, I've been having the exact same problem as you; almost weak, whispy growth in all my plates lately.

Before reading this thread, I was convinced my nutes were too high, and tried A BUNCH of recipes both as-is and with 25% less nutes. No good results so far.
I tried Potato flake + honey, Liquid malt Extract (alas, no dry extract here), diluted grain soak water, diluted grain boil water, and BRF.

I'll up the nutes a bit like Benson says in my next agar batch and let you know if the situation improves.

Good luck man, I understand your frustration with not getting the results you want from agar. We'll make it through :evil:


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Offlinemoshe1

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Re: Help me understand my agar [Re: jomanda1990]
    #27327189 - 05/28/21 10:12 PM (17 days, 5 hours ago)

my main concern is identifying whats going on when i look at it! having problems is one thing not being able to identify them is a lot worse lmao

but i would take weak growth over mold any day- cause apparently all my trays have contam by the feedback im getting


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Edited by moshe1 (06/02/21 05:26 AM)


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InvisibleFeasoghorm

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Re: Help me understand my agar [Re: moshe1]
    #27327239 - 05/28/21 11:07 PM (17 days, 4 hours ago)

Yeah dude make some plates with more nuteage and transfer to those plates from the crazy looking ones to see if it comes out nice.


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OfflineCrackatoaS
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Re: Help me understand my agar [Re: Feasoghorm] * 1
    #27327280 - 05/28/21 11:58 PM (17 days, 3 hours ago)

My recipe is 10g agar, 7-8g LME per 600ml plus nutritional yeast. The yeast has really helped.


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Offlinemoshe1

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Re: Help me understand my agar [Re: Crackatoa]
    #27327330 - 05/29/21 12:54 AM (17 days, 2 hours ago)

thanks crackatoa, ill try adding some yeast and see what it does. Congrats on your TC btw

would you say that its still possible that i'm recipe/nutritional issues only?  or you are also pretty made up as well that my plates all having a contam issue going on


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Edited by moshe1 (05/29/21 12:55 AM)


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InvisibleFeasoghorm

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Re: Help me understand my agar [Re: moshe1]
    #27327334 - 05/29/21 01:04 AM (17 days, 2 hours ago)

I understand that you did not ask me, but at this point im basically convinced that what we are seeing is a nute issue. I feel bad that i may have put the contamination theory in your head. I shud knw this shit by now, but i was basically retarded till the beginning of this year. So you'll have to excuse me.


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Offlinemoshe1

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Re: Help me understand my agar [Re: Feasoghorm]
    #27327352 - 05/29/21 01:33 AM (17 days, 2 hours ago)

no problems my dude, and i appreciate hearing that from anyone actually, so i'll take it. Seriously, go search for an answer to an issue like im having with the search function you ll end up ripping your hair out.

you will go from being convinced its contam, to convinced its nutrients and back and forth with every other thread you read into lol, i'm definitely reporting back step by step to put a nail in this coffin


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Edited by moshe1 (05/29/21 01:34 AM)


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OfflineBlackrainbow2
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Re: Help me understand my agar [Re: moshe1]
    #27327372 - 05/29/21 01:49 AM (17 days, 2 hours ago)

another option is get your self a few plates from a vendor and see if they preform normal and that may get you answers, on if it's your agar or something else.. I think 10 plats are like $22..


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Re: Help me understand my agar [Re: Blackrainbow2]
    #27327686 - 05/29/21 10:34 AM (16 days, 17 hours ago)

I believe it may be both, I feel like there's still some contamination in there hiding they can cleaned and some have really good looking edges to snag but I also feel once we figure this nute issue out the good myc will be easy to see and snag.


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Offlinemoshe1

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Re: Help me understand my agar [Re: Crackatoa]
    #27328174 - 05/29/21 06:27 PM (16 days, 9 hours ago)

bumpdate :

fresh melmak spores went to agar just over a week ago. went a day too long but gonna transfer out of these today. showing some similarities to previous plates though- same agar recipe.  i'll be transferring out of the odd looking plates from the original post next couple days with a higher nutritional agar and lower to see the difference




--------------------


Edited by moshe1 (05/29/21 07:38 PM)


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OfflineCrackatoaS
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Re: Help me understand my agar [Re: moshe1]
    #27328193 - 05/29/21 06:47 PM (16 days, 9 hours ago)

:thisisgonnabegood:


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Offlineverum subsequentis
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Re: Help me understand my agar [Re: Crackatoa]
    #27328239 - 05/29/21 07:33 PM (16 days, 8 hours ago)

Looking good. Looks like there's goodies in all those plates.


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Offlinemoshe1

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Re: Help me understand my agar [Re: verum subsequentis]
    #27328242 - 05/29/21 07:37 PM (16 days, 8 hours ago)

room still scrubbing- im thinking i should do the transfers in my SAB this time around? or should i continue working in front of flow?  haven't used the good ol' glove box in so long


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Offlineverum subsequentis
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Re: Help me understand my agar [Re: moshe1] * 1
    #27328245 - 05/29/21 07:39 PM (16 days, 8 hours ago)

Why wouldn't you use the flow? I would.


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Offlinemoshe1

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Re: Help me understand my agar [Re: verum subsequentis]
    #27328250 - 05/29/21 07:43 PM (16 days, 8 hours ago)

idk, i was under the impression transferring from contam plates infront of the flowhood = bad

i'd much much rather keep working in front of the hood lol spent a lot of time building the big boy


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Offlineverum subsequentis
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Re: Help me understand my agar [Re: moshe1] * 1
    #27328254 - 05/29/21 07:45 PM (16 days, 8 hours ago)

It's only bad if you are opening a plate with nasties that are sporulating. Even then, I've pulled it off in the flow several times.


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Re: Help me understand my agar [Re: verum subsequentis]
    #27328272 - 05/29/21 08:00 PM (16 days, 7 hours ago)

Sector n xfer only your strongest growth. Sacrifice the rest; not ballin. Shooting for rhizmophoric action. If you went from spores id xfer 2 or 3 times before noccing up spawn at the least.


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Offlineverum subsequentis
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Re: Help me understand my agar [Re: ConcreteWaves]
    #27328362 - 05/29/21 09:16 PM (16 days, 6 hours ago)

Shooting for rhizo seems to be somewhat of a fool's errand. Shoot for clean plates. I believe I'd have perfectly fine transfers on the next round if those were mine.


Edited by verum subsequentis (05/29/21 09:16 PM)


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InvisibleFeasoghorm

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Re: Help me understand my agar [Re: verum subsequentis] * 1
    #27328445 - 05/29/21 10:24 PM (16 days, 5 hours ago)

Them are some pro germ plates :thumbup:

Cant wait to see your findings about the plate nutes. Your doing the Lord's work, moshe1


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Offlinemoshe1

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Re: Help me understand my agar [Re: Feasoghorm]
    #27328472 - 05/29/21 10:45 PM (16 days, 5 hours ago)

Quote:

verum subsequentis said:
Shooting for rhizo seems to be somewhat of a fool's errand. Shoot for clean plates. I believe I'd have perfectly fine transfers on the next round if those were mine.




hoping the first transfers come out clean as well, lets see what happens. just finished up with another set of 20 plates, most of em were used on those new germs, and a few were used on those ugly t1 and t2s from first post.


Quote:

Feasoghorm said:
Them are some pro germ plates :thumbup:

Cant wait to see your findings about the plate nutes. Your doing the Lord's work, moshe1




thanks! same here man, hoping to save time for all the future people on this. my germ plates looked pretty decent on my first sets as well before they went wild on me, fingers crossed.


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Offlinemoshe1

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Re: Help me understand my agar [Re: moshe1]
    #27330703 - 05/31/21 09:30 PM (14 days, 6 hours ago)

BUMPDATE  5/31

Here are those fresh melmaks on their first transfer today. These are also 10g agar to 7-8g LME to just over 400 ML water.

Prepped two new agar bottles, one with ~5G LME, 10G Agar, 400ML and the other with the ~7G LME + 1G Nutritional Yeast, 400ML.  Will transfer to those next coming days.

Just got back from the supermarket and they had very fine stoneground BRF on clearance half off, snagged 5 pounds for only like 10 bucks- so at some point BRF agar will be on trial






transparent layer of myc growing outwards only visible well under light hard to take pictures.


Edited by moshe1 (06/01/21 03:33 AM)


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Offlineverum subsequentis
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Re: Help me understand my agar [Re: moshe1]
    #27330708 - 05/31/21 09:37 PM (14 days, 6 hours ago)

Let's see how they grow out. I think you're lookin pretty good.


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Re: Help me understand my agar [Re: verum subsequentis]
    #27330735 - 05/31/21 10:04 PM (14 days, 5 hours ago)

I've also been having the exact same problem, put in yeast and it's like night and day. I'll post pics when I have wifi set up


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Offlinemoshe1

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Re: Help me understand my agar [Re: jomanda1990]
    #27330926 - 06/01/21 02:14 AM (14 days, 1 hour ago)

Quote:

Ashtray161 said:
I've also been having the exact same problem, put in yeast and it's like night and day. I'll post pics when I have wifi set up




hey which problem are you talking about because im trying to figure a few out.

One is the "weak", thin and transparent myc instead of thicker myc- ruled out genetics because its happening on all of my varieties. Weak in quotes because its not the myc fault.

Second problem is figuring out what is contam and what isnt, because the white fuzzy/cottony stuff is easy to confuse for tementose (at least for me) and it stays white with no discoloration. Other contams are very obvious

I prepped some agar, added a gram of yeast to the recipe for one of the bottles- so lets see if it does anything for my plates like yours


Edited by moshe1 (06/02/21 09:36 PM)


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Offlinemoshe1

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Re: Help me understand my agar [Re: moshe1]
    #27332878 - 06/02/21 06:30 PM (12 days, 9 hours ago)

BUMPDATE 06/02:

here are melmak transfer 1s + close ups. gonna be out of town for the next 4 days- until monday. Wondering if i should leave them or transfer out again before i head out.
To test nutes im gonna bust open the melmak germ plates again before they're all grown out and get some new T1s on the high/low nutrient dishes as well to see if they look any different to these.  Also gonna transfer some of those older plates with the weak growths and contams to them and see how they react. some of the older plates i'll be using are pictured last.


MAKS



MAK CLOSEUPS







PRIOR PLATES

To test the high/low nute plates ill also be transferring out of these that appear contamd and/or super thin and transparent, BBM, MAZ, PE6 and LGT (shown underside cause of condensation)



Edited by moshe1 (06/02/21 08:23 PM)


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InvisibleFeasoghorm

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Re: Help me understand my agar [Re: moshe1]
    #27333046 - 06/02/21 09:05 PM (12 days, 6 hours ago)

One of my Mak plates looks exactly like plate#2 of the Mak close-ups.
:notsureif:
Cant tell if it's mold or sectoring.


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Offlinemoshe1

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Re: Help me understand my agar [Re: Feasoghorm]
    #27333271 - 06/03/21 02:32 AM (12 days, 1 hour ago)



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Re: Help me understand my agar [Re: moshe1]
    #27333274 - 06/03/21 02:38 AM (12 days, 1 hour ago)

I have a Cambodian that grows like that, always looks like contamination but never shows anything when transferred away from the dish.


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Offlinemoshe1

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Re: Help me understand my agar [Re: DERRAYLD]
    #27339338 - 06/07/21 08:55 PM (7 days, 6 hours ago)

BUMPDATE 06/07


The GREEN agar has 1G Yeast to 7G LME  and the BLUE agar has only 5G LME
not much difference but the lower nutritional agar (blue) moving a little slower. Still have super fuzzy growth happening


PE6
more thin and weak looking growth on the two test plates


Edited by moshe1 (06/10/21 04:52 PM)


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Offlinemoshe1

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Re: Help me understand my agar [Re: moshe1]
    #27343195 - 06/10/21 04:15 PM (4 days, 11 hours ago)

BUMPDATE - 06/10

Today i am preparing another batch of AGAR, one BRF + YEAST and another LME + YEAST to see how the BRF does as well.

below are a couple of the different varieties i have and their reactions to the two types of agar i put them on.

BLUE AGAR = 5G LME 10G AGAR   
GREEN AGAR = 7G LME 1G YEAST 10G AGAR



Also, those melmak transfer 1s that i didn't touch yet, would any of you feel confident putting these to grain?



--------------------


Edited by moshe1 (06/11/21 04:04 AM)


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OfflineChocyMilky
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Re: Help me understand my agar [Re: moshe1]
    #27343604 - 06/10/21 10:10 PM (4 days, 5 hours ago)

Quote:

moshe1 said:
Also, those melmak transfer 1s that i didn't touch yet, would any of you feel confident putting these to grain?






That first one and the last two of the second pic looking nice for sure. I like to see perfect contiguous growth, and any disruptions to growth I like to examine by looking through the agar plate with a strong light behind it and checking for any obvious signs of contamination, bacteria rounds, etc.

It looks like you nailed it though with determining the nutrient deficiency (possible competing fungi too..).., Ive definitely experienced something similar.


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Re: Help me understand my agar [Re: ChocyMilky] * 1
    #27343670 - 06/10/21 11:27 PM (4 days, 4 hours ago)

DUDE I've been having the exact same problem. I've tried like 7-8 spores syringes and I can't get the nice thick rhizomorphic mycelium. Last year I pulled off a 30 tub grow clean, but I mistakenly threw away all my cultures intending to start over. I've made at least 100 plates recently and they all look pretty much the same as yours. Many show very faint, tomentose growth that extends/grows way faster than the thicker myc. Really hoping the nute thing is the solution. Out of the 50 plates I made last week LITERALLY ONE has nice looking growth and I don't trust it. I'm very glad you made this thread. I was about to make one myself.

I'm seriously starting to wonder if syringes bought from popular vendors are contaminated with a mycovirus or something, not likely but I can't explain this shit any other way. Been growing for 6 years and never seen anything this fucking puzzling.


--------------------
 


Edited by afrekcan (06/10/21 11:31 PM)


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Offlinemoshe1

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Re: Help me understand my agar [Re: afrekcan]
    #27344742 - 06/11/21 08:01 PM (3 days, 7 hours ago)

Afrekcan, of all the things i had to figure out with this hobby so far this is the most puzzling for me as well, next to some fruiting issues i've experienced.

so far i find that the 5G LME agar seems to do more rhizo growth, even though the fuzzy shit is still in most plates. Of all the plates i've inoculated and transferred across 6 varieties so far only a handful have looked good, and that wasn't even til just very recently. Everything else still is fuzzy and has major inconsistencies.

i'll keep updating as I go. I recently found a rogue plate that is growing just white fuzzy shit, i never inoculated it. I'm assuming whatever is growing on it is the culprit- so far everything is loosely based on very little evidence i've gathered though, so cant be conclusive just yet.

These are across different vendors and trades too, so shit got me major stumped- you're not alone!

Quote:

ChocyMilky said:

It looks like you nailed it though with determining the nutrient deficiency (possible competing fungi too..).., Ive definitely experienced something similar.




i think its actually too much nutrition and not deficiency. Cant say for sure though, as you can see in the sample size pic of my plates above, BBM and MAK plates goin rhizo on the low nute plates, but PE and MAZ barely showing any color on them.  It could also still be too much nutrition + a very hard to nail contam-


heres a contam plate that hasn't been exposed to any mushy spores- so whatever that is made its way to some of the plates perhaps? Much much darker under light compared to myc







--------------------


Edited by moshe1 (06/11/21 11:45 PM)


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OfflineChocyMilky
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Re: Help me understand my agar [Re: moshe1]
    #27345749 - 06/12/21 06:03 PM (2 days, 9 hours ago)

When I last experienced this it was due to using old (and i guess expired) Malt Extract Agar (MEA) pre-formulated from a trusted vendor. I had last used it about 2 months prior following the same recipe with normal success.

So it may be that your ingredients have expired..or what I am really saying to those reading is that expired ingredients may be to blame. Once I bought a fresh batch of MEA from same trusted vendor, I once again had the growth one would expect from the same MSS syringes.

Now I keep it vac sealed between uses., maybe that will extend the shelf life.., maybe refrigeration would be good too, I'm not sure.


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Offlinemoshe1

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Re: Help me understand my agar [Re: ChocyMilky]
    #27345779 - 06/12/21 06:35 PM (2 days, 9 hours ago)

https://www.amazon.com/gp/product/B0038O5N4I/ref=ppx_yo_dt_b_search_asin_title?ie=UTF8&psc=1

Basically bought this back in late February. Been using it since, but not sealed just in the bag with a hole i cut out on the top in the cupboard


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Re: Help me understand my agar [Re: moshe1]
    #27347032 - 06/13/21 07:18 PM (1 day, 8 hours ago)

BUMPDATE 06/13

Waiting for these mak t1s to finish filling up the plate, i think im going to take them straight to grain and hope for a good clone to grab. Trying to figure out how what path im going to take for fruiting.



two mak plates, one on 5G LME (blue) and the other on 7G LME + 1G YEAST (green). I have other plates that are mixed but this shows the first signs of rhizo growth on this batch. very fuzzy / tementose in plates though, still possibly contam?



my best looking plate in terms of rhizo growth across all my dishes so far. BBM that came from one of my first plates i started with.



more of those BBM plates im still transferring. Some are starting to look clean i think.



a BBM on low nute plate thats still throwing a lot of fuzzy, so again im pointing out the best examples



the xfers i am going to take from today. There is one mak plate in there with the best looking rhizo i have from that variety. They will be going to high nute BRF agar and LME agar.  Going to prep some 5G LME today to transfer next since its showing to grow the most rhizos im seeing on my plates so far. Hopefully things are looking (mostly) clean here



heres that contam plate today, i never put any spores on these. that white stuff i think may be whats fucking my plates



luckily i took a clone sample from a MS PEU project that was failing, and it through out pins in just 4 weeks or so, so going to use this to start fruiting these until im consistent with my AGAR candidates





Edited by moshe1 (06/13/21 07:26 PM)


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Re: Help me understand my agar [Re: moshe1] * 1
    #27347123 - 06/13/21 08:44 PM (1 day, 7 hours ago)

Thanks for posting this as well as updating.

Cleaning up germination plates have been a complicated process for me.


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Offlinemoshe1

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Re: Help me understand my agar [Re: Atomsplit]
    #27347134 - 06/13/21 08:54 PM (1 day, 6 hours ago)

that BBM plate with all the rhizo growth came from the weird looking plates i posted about initially that got me the advice to just start over with fresh germs, now its the first plate to look like a proper agar dish by T4- so just keep going


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Edited by moshe1 (06/14/21 03:01 AM)


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