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esse_jeremy
the deconstructor


Registered: 05/28/16
Posts: 173
Loc: Italy
Last seen: 1 year, 3 months
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d(r)ied print???
#23419742 - 07/07/16 10:08 AM (7 years, 6 months ago) |
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I made several print to agar attempts with this procedure:
-half the agar recipe to a soft surface
-less nutes
-sterelized for 30 min in pc (no pour tek, litle hole with micropore tape as filter)
-cool down and place in a SAB
-use a loop (red hot, cooled in the reciving plates, grab spore, tuch the plates)
-store the innocced plates in a dark and everyday cleaned place
now, before I bought my SAB I got a contam after 3 days I innoc. them (so I assume I sterelized them properly but I fail during innoc)
after several tries, i bought a SAB and inoocc. two plates
the problem is: after a 1.5 week there's nothing growing on the plate.....ok I got no contam so the SAB worked well...but I also have no MYC....nothing.
I'm scared to death that the print is dead or dry...but the agar should reidrate the spore......so why they don't germ??
I did something wrong with my loop? I have to put a drop of sterile water on the print??
I'm afraid...this is my 6th attempt and I never had a myc (also I spend 3 hours a day reading post on the forum to understand the theory and improve but the only thing I saw are people having amazing grow with my same tek....and sometimes also with less gear or with improper sterile tek)
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things does not exist, everything is a process, so we proceed
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mupetmower
Mower of Muppets



Registered: 03/29/16
Posts: 3,036
Loc: Here and There
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It could be the spores. I mean sometimes they can just have a hard time germinating. Just keep trying. Keep them for at least 3 weeks to make sure it's not jaunt taking a while for them. I've had a couple plates not grow for about 3 weeks and then when I was about to throw them out I noticed they had started germinating finally.
After about a month I would throw them out, though. But just keep trying. You should get some germination as long as the print isn't super old.
-------------------- -The wise man never stops seeking knowledge.
-I wanna feel the change consume me, feel the outside turning in. I wanna feel the metamorphosis and cleansing I've endured within.
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weetsie
unlicensed tub surgeon



Registered: 05/08/11
Posts: 572
Loc: United Kingdom
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Is there a visible amount of spores on the agar?
A dry print is ok, they still work, just take longer and occasionally stick like a fucker to the foil making it hard to transfer them.
You can try making a spore syringe or digging a little hole in the agar with the loop and putting a drop of water in it with the spores.
Old spores can take a while, 3+ weeks and beyond a certain point it really becomes a numbers game. If you're not noticing contams you can increase your odds by adding spores to multiple sites per plate, I do about 5 or 6 with stubborn spores and even then it sometimes takes 3 plates and a month before you get any that agree to germinate.
-------------------- Active grow logs: Oysters on Straw Pellets Trade list
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esse_jeremy
the deconstructor


Registered: 05/28/16
Posts: 173
Loc: Italy
Last seen: 1 year, 3 months
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Re: d(r)ied print??? [Re: weetsie]
#23419833 - 07/07/16 10:40 AM (7 years, 6 months ago) |
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I think I can experiment adding spore to one of the plates
Also yes there is a visible ammount (not clearly visible but if u get closer u can see some spore).
And what are pros and cons of making several sites per plates??
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things does not exist, everything is a process, so we proceed
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impatientguy
Ganjalf a very mighty lab wizard



Registered: 11/26/14
Posts: 5,054
Loc: USA
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You should hydrate the spores IMO
They will germinate much more readily that way.
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esse_jeremy
the deconstructor


Registered: 05/28/16
Posts: 173
Loc: Italy
Last seen: 1 year, 3 months
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Quote:
impatientguy said: You should hydrate the spores IMO
They will germinate much more readily that way.
I'm afraid that if I open them again I will have a contam....I don't know if it's worth 'cause lot of ppl said that this will not speed up germination....I'm so confused
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things does not exist, everything is a process, so we proceed
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Mad Season
hookers and blackjack



Registered: 09/16/12
Posts: 12,666
Loc: Canada
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You can also drop the agar a bit to make it more watery, and up the nutrients. Lower nutrients will be harder to germinate
1.5g of agar and 2g of malt extract would be awesome for germinating.
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dankington
The Stranger




Registered: 03/14/15
Posts: 4,577
Loc: 8te
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Quote:
Mad Season said: You can also drop the agar a bit to make it more watery, and up the nutrients. Lower nutrients will be harder to germinate
1.5g of agar and 2g of malt extract would be awesome for germinating.
you might mention the 200ml or so of water.
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Mad Season
hookers and blackjack



Registered: 09/16/12
Posts: 12,666
Loc: Canada
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Lol woops. Actually per 100ml
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dankington
The Stranger




Registered: 03/14/15
Posts: 4,577
Loc: 8te
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oooh, okay. yeah. it get tricky with agar ratios sometimes.  I wasn't sure where you were directing him. But yeah, 100-120ml of water for 1.5g agar 2g ME sounds pretty stellar for spore germination.
especially being I'm in the US, so more used to other units of measure. maybe I should move back to canada lol.
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esse_jeremy
the deconstructor


Registered: 05/28/16
Posts: 173
Loc: Italy
Last seen: 1 year, 3 months
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Quote:
dankington said: oooh, okay. yeah. it get tricky with agar ratios sometimes.  I wasn't sure where you were directing him. But yeah, 100-120ml of water for 1.5g agar 2g ME sounds pretty stellar for spore germination.
especially being I'm in the US, so more used to other units of measure. maybe I should move back to canada lol.
is the exact ammount of agar-nutes-water that I used for my plates 1.5% agar 2% nutes
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things does not exist, everything is a process, so we proceed
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Grey
⇜ ✯ ⇝



Registered: 11/06/14
Posts: 6,223
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Is there a date on your print? Did you buy it?
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AMU Q&A If you don't have a plan of your own, you'll become a part of somebody else's.
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esse_jeremy
the deconstructor


Registered: 05/28/16
Posts: 173
Loc: Italy
Last seen: 1 year, 3 months
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Re: d(r)ied print??? [Re: Grey]
#23422753 - 07/08/16 08:34 AM (7 years, 6 months ago) |
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I bought it but there's no date, the vendor is a sponsor of the forum...and is professional...I don't think they sell me dead print, I hope!!!!
also I storing them upside down after innoc. can this affect the GE and the grow of myc??
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things does not exist, everything is a process, so we proceed
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Grey
⇜ ✯ ⇝



Registered: 11/06/14
Posts: 6,223
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Okay. I don't think they would either.
No, storing upside down is fine. Sometimes they just take some time to germinate.
After you cool the loop and scrape up spores, do you swipe over the spot you cooled your loop? It's usually a little watery where you cool the loop/blade and is helpful for Germination.
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AMU Q&A If you don't have a plan of your own, you'll become a part of somebody else's.
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esse_jeremy
the deconstructor


Registered: 05/28/16
Posts: 173
Loc: Italy
Last seen: 1 year, 3 months
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Re: d(r)ied print??? [Re: Grey]
#23422825 - 07/08/16 09:08 AM (7 years, 6 months ago) |
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Yep, I've done it, it's possible that the spore remain on the loop and didn't stick to agar??(just my paranoid)
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things does not exist, everything is a process, so we proceed
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Grey
⇜ ✯ ⇝



Registered: 11/06/14
Posts: 6,223
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Possible, but the spores are very tiny so you may have still swiped some.
I think someone suggested it, but instead of just swiping across the surface try stabbing or "cutting" the agar. Basically drag your loop through the agar.
If your worried they're old or dry, maybe check this thread out.
https://www.shroomery.org/forums/showflat.php/Number/15383615
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AMU Q&A If you don't have a plan of your own, you'll become a part of somebody else's.
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esse_jeremy
the deconstructor


Registered: 05/28/16
Posts: 173
Loc: Italy
Last seen: 1 year, 3 months
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Re: d(r)ied print??? [Re: Grey]
#23423369 - 07/08/16 11:55 AM (7 years, 6 months ago) |
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Ty for the info man!!!!
Just one more question, where can i retrive the stopper for the syringe?? I never heard about it, is there a way to do that with similar obj??
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things does not exist, everything is a process, so we proceed
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Grey
⇜ ✯ ⇝



Registered: 11/06/14
Posts: 6,223
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Just about any of our sponsors would carry them. Just make sure they're autoclavable. I can pm you a link if you need.
I'm not sure of another way to plug a syringe for that process.
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AMU Q&A If you don't have a plan of your own, you'll become a part of somebody else's.
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Munchauzen


Registered: 06/22/11
Posts: 14,342
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Quote:
esse_jeremy said: Ty for the info man!!!!
Just one more question, where can i retrive the stopper for the syringe?? I never heard about it, is there a way to do that with similar obj??
No need for a stopper or rubber band. Idk why stro thought that was necessary, because they will hydrate just by being in the syringe.
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Mad Season
hookers and blackjack



Registered: 09/16/12
Posts: 12,666
Loc: Canada
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The prints are how old? I really doubt them being dry is the problem here unless they're at least 2 years old.. 1.5 weeks is a bit long, but not like OMFG these spores are unviable!
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