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Offlinejee3apha
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First attempt at PF-tek: no mycelium after 2 weeks * 1
    #27840927 - 06/28/22 04:34 PM (2 years, 6 months ago)

So after studying countless guides and posts about this topic, I decided to give it a try as soon as the temperature was high enough where I live. After 2 weeks since inoculation, out of 6 jars (4 inoculation points each) only one (and in one point only) seems to have generated some mycelium. Provided it's not just contamination...



I tried to follow the tek closely, but here are the main key points:

  • spores was P. Cubensis (Amazonian) in a syringe, kept in the fridge as soon as I got it
  • substrate with 2/1/1 of verm/water/BRF respectively
  • added the top layer of dry verm
  • jars have been sterilized in a regular pot for 3 hours then let them cool down for about 18 hours
  • inoculation in SAB, as per tek, needle red hod, then cooled, then inoculated a few drops toward the glass
  • syringe flamed once per jar using the gas stove flame
  • jars left germinating in a closet inside a cardboard shoe box with nothing to obstruct the inoculation holes of the jars, 82-86°F (28-30°C) and 50-60% RH (as seen in the photo, there's plenty of moisture in the jars, I think)


I'm unsure about the following:

  • the syringe wasn't good, but there's no easy way for me to figure this out I guess...
  • the BRF I used wasn't good
  • I didn't press the substrate thus there are some too-airy spots, and the spores didn't find the proper conditions/nutrients, or the way to move freely across the substrate


Out of the above points only the first two might justify this complete failure. So I'm unsure about what to try next, should I wait for another couple of weeks? But then why would there be that outlier in the photo? Should I throw everything away and start over? Fact is that I'd do pretty much the same things, except for maybe try to compress the substrate a bit more, but I mean, I would've expect something in the range of 8-10 successful inoculation points if that was the case, not only one.

Thoughts on this? I hope I've provided all the necessary information, if not, feel free to ask.

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Invisiblesolarentity9
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Registered: 06/27/22
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Re: First attempt at PF-tek: no mycelium after 2 weeks [Re: jee3apha] * 1
    #27840961 - 06/28/22 04:53 PM (2 years, 6 months ago)

imo your substrate looks weird.

Heres how I make PF-tek Substrate


Vermiculite 2 parts
Water 1 part
Mix vermiculite and water then STRAIN the excess water from verm.

Now, add 1 part Brown rice flower, spoon into jar, dont pack it down. then wipe jar rim and add dry substrate layer, etc.


from what i can see in the photo:
ur substrate is clumpy
ur substrate is too wet, and the vermiculite i can see isnt coated in vermiculite.


id start a new grow, read the tek more carefully next time.

Edited by solarentity9 (06/28/22 04:56 PM)

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OfflineThe Tao
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Re: First attempt at PF-tek: no mycelium after 2 weeks [Re: jee3apha] * 3
    #27841005 - 06/28/22 05:11 PM (2 years, 6 months ago)


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OfflineKROM
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Re: First attempt at PF-tek: no mycelium after 2 weeks [Re: solarentity9]
    #27841013 - 06/28/22 05:13 PM (2 years, 6 months ago)

Shot in the dark here, but did you vigorously shake the syringe before using it to distribute the spores throughout the liquid? Sometimes they clump together. Also, 80+ is a little hot, cubensis prefers 65-75f. And was the box to block light? Cubensis benefits from lighting at all life stages.


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Invisiblesolarentity9
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Re: First attempt at PF-tek: no mycelium after 2 weeks [Re: KROM]
    #27841376 - 06/28/22 08:45 PM (2 years, 6 months ago)

That substrate isn't correct.

Soaked/Wet/Water-Absorbed Vermiculite should then have Brown rice flour stuck to the wet verm. I just see wet verm here, missing the BRF proper coating.

The clumps are unnatural.
It looks too wet.

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OfflineVibetyme
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Re: First attempt at PF-tek: no mycelium after 2 weeks [Re: jee3apha]
    #27841399 - 06/28/22 08:55 PM (2 years, 6 months ago)

Don't throw them away.  Just be patient. 

I have had several jars not show growth until the second or third week. 

Yeah it looks a little funny but I bet at least two or three of your jars colonize. Hopefully. 

What do the other jars look like?


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OfflineLogicaL ChaosM
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Re: First attempt at PF-tek: no mycelium after 2 weeks [Re: jee3apha]
    #27841719 - 06/29/22 12:33 AM (2 years, 6 months ago)

You made your substrate too wet (and too loose, but thats a minor issue). You can try put them in the sunlight and hope the water evaporates away but I would start over thou.


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OfflineSingularFusion
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Re: First attempt at PF-tek: no mycelium after 2 weeks [Re: LogicaL Chaos] * 1
    #27841725 - 06/29/22 12:48 AM (2 years, 6 months ago)

when I used to do PF I always had best results making the mix in a large mixing bowl or salad bowl

I would add verm and water together in the bowl and mix thoroughly, there will be no excess to pour off, the verm alone will hold all the water IME

then I take the rice flour and put into a sieve and sieve it slowly onto the wet mixture that is nicely spread out in the bowl, while pausing the sieving to mix the bowl gently with something that keeps the mix light, you want a whisk or a fork, not a big paddle or spoon or spatula. Slowly keep sieving the flour in and mixing until it's all in. It should be entirely homogenous

spoon into jars, wipe the rims and then add dry verm layer

I dunno where you went wrong OP but it looks a little wrong as the others have said already, however you may get lucky, the PF tek is extraordinarily resilient

I would start some more things anyway

all the best with it :rockon:

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Offlinejee3apha
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Re: First attempt at PF-tek: no mycelium after 2 weeks [Re: solarentity9]
    #27841760 - 06/29/22 02:30 AM (2 years, 6 months ago)

I agree that's too clumpy, I was afraid to press it too much so I just "let it fall" into the jar, hence causing the clumps and the airy spots. That's definitely a mistake and something that I learned the hard way.

I did pretty much as you said, including wiping the rim, the only thing is that I mixed in the wrong order (a stupid mistake I realized right away...), that is, verm, rice, then water. The amount of water looked right though, I respected the 2/1/1 ratio, also I tried to squeeze the thing and only a few drops did fall, I remember I saw this in the Updated PF-Tek Guide 2017 Edition, unless I misunderstood the instructions.

Quote:

and the vermiculite i can see isnt coated in vermiculite.




What do you mean by that?

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Offlinejee3apha
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Re: First attempt at PF-tek: no mycelium after 2 weeks [Re: KROM]
    #27841762 - 06/29/22 02:34 AM (2 years, 6 months ago)

Quote:

KROM said:
Shot in the dark here, but did you vigorously shake the syringe before using it to distribute the spores throughout the liquid? Sometimes they clump together. Also, 80  is a little hot, cubensis prefers 65-75f. And was the box to block light? Cubensis benefits from lighting at all life stages.




I did shake it yeah.

Yes they sat still in the dark, I can see that the light thing at this stage is a pretty controversial topic, I couldn't figure out who was right so I stick with the original tek. I guess I could just put them under light now, I mean before throwing everything away...

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Offlinejee3apha
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Re: First attempt at PF-tek: no mycelium after 2 weeks [Re: Vibetyme]
    #27841764 - 06/29/22 02:35 AM (2 years, 6 months ago)

Quote:

Vibetyme said:
What do the other jars look like?




Exactly like the one in the photo, just without any sign of mycelium.

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Offlinejee3apha
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Re: First attempt at PF-tek: no mycelium after 2 weeks [Re: LogicaL Chaos]
    #27841765 - 06/29/22 02:37 AM (2 years, 6 months ago)

That's definitely something I could try before throwing everything away, hopefully I would learn something from it!

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Offlinejee3apha
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Re: First attempt at PF-tek: no mycelium after 2 weeks [Re: SingularFusion]
    #27841767 - 06/29/22 02:42 AM (2 years, 6 months ago)

Quote:

Nef said:
when I used to do PF I always had best results making the mix in a large mixing bowl or salad bowl

I would add verm and water together in the bowl and mix thoroughly, there will be no excess to pour off, the verm alone will hold all the water IME

then I take the rice flour and put into a sieve and sieve it slowly onto the wet mixture that is nicely spread out in the bowl, while pausing the sieving to mix the bowl gently with something that keeps the mix light, you want a whisk or a fork, not a big paddle or spoon or spatula. Slowly keep sieving the flour in and mixing until it's all in. It should be entirely homogenous

spoon into jars, wipe the rims and then add dry verm layer

I dunno where you went wrong OP but it looks a little wrong as the others have said already, however you may get lucky, the PF tek is extraordinarily resilient

I would start some more things anyway

all the best with it :rockon:




Thank you mate! That's a nice piece of advice about how to get the substrate right.

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Offlinejee3apha
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Re: First attempt at PF-tek: no mycelium after 2 weeks [Re: jee3apha]
    #27841771 - 06/29/22 02:53 AM (2 years, 6 months ago)

I want to thank you all of you guys for all the precious tips and insights.

So, it looks like that I just fucked up the substrate part, in at least two ways:

  • by mixing the components in the wrong order (verm -> BRF -> water), with the BRF not sticking properly to the verm stones
  • by not gently pressing/adjusting the substrate in the jar so to avoid airy spots


I guess I learned something new, love you guys!
Will keep you posted so to hopefully help others in the same situation.

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OfflineRotnpins
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Re: First attempt at PF-tek: no mycelium after 2 weeks [Re: jee3apha] * 2
    #27841776 - 06/29/22 03:04 AM (2 years, 6 months ago)

I had a batch of pf tek jars that were overly wet and looked similar... they all ended up colonizing anyway.. I would definitely wait it out a while before tossing them

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Offlinejee3apha
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Re: First attempt at PF-tek: no mycelium after 2 weeks [Re: jee3apha]
    #27867136 - 07/18/22 06:31 AM (2 years, 5 months ago)

So a little update. I ended up tossing everything away and starting over using the remaining half-ish syringe I had. This time I tried to fix all the above errors. Today, after 20 days there's is no mycelium at all. At this point I guess it's safe to assume that the syringe wasn't right...

I have a follow-up question though, in both cases when I opened the jars to throw away the substrate I noticed some peculiar smell, nothing that would resemble mold or anything related, but sill not pleasant. It's a sweetish smell that loosely resembles something between warm cheese and pickled cucumber (I'm afraid my English is not good enough to give you a better description). There's no visible contamination, at least to my inexperienced eye, and all the jars have it.

I guess it's normal that the substrate has some kind of smell after 20 days sitting in a moist jar with some organic material?

Does this description sound familiar? Is this a common hallmark of some contamination?

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OfflineRacoon4
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Re: First attempt at PF-tek: no mycelium after 2 weeks [Re: jee3apha]
    #27867248 - 07/18/22 08:33 AM (2 years, 5 months ago)

likely some bacteria colonized the germination site preventing myc from germinating, judging from your description of the smell. Your syringe is too dirty, theres nothing you can do about that except try your best to grow a fruit so you can make your own spore print to refill the syringe.

- I don't know if your doing this yet but use the tiniest drop you possibly can squirt out of the syringe. The less spores you use, the less contamination in your jars so your chances of success are actually increased.
- The drier your cake is, the better. Drier cakes are more resistant to contam growth, but myc can colonize drier cakes just fine. A dry cake wont really effect germination because the water from the syringe will provide plenty. Even if its just a tiny drop. Your cakes look really wet, more moisture than ive ever had in a cake.

Edited by Racoon4 (07/18/22 08:35 AM)

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Offlinejee3apha
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Re: First attempt at PF-tek: no mycelium after 2 weeks [Re: Racoon4]
    #27867320 - 07/18/22 09:29 AM (2 years, 5 months ago)

Quote:

Racoon4 said:
likely some bacteria colonized the germination site preventing myc from germinating, judging from your description of the smell. Your syringe is too dirty, theres nothing you can do about that except try your best to grow a fruit so you can make your own spore print to refill the syringe.




Thanks for confirming this.

Quote:

- I don't know if your doing this yet but use the tiniest drop you possibly can squirt out of the syringe. The less spores you use, the less contamination in your jars so your chances of success are actually increased.
- The drier your cake is, the better. Drier cakes are more resistant to contam growth, but myc can colonize drier cakes just fine. A dry cake wont really effect germination because the water from the syringe will provide plenty. Even if its just a tiny drop. Your cakes look really wet, more moisture than ive ever had in a cake.




Those are two good pieces of advice. Actually the above photo was from my first attempt, here's the second one:



This time I tried to fix the previous errors. But I definitely might have inoculated with too much liquid...

Also I wonder, is it OK (assuming that the syringe is initially viable) to reuse for a second batch the remaining liquid? Especially if only a few drops are used, a whole syringe is way too much to colonize say 5 o 6 jars only.

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OfflineRacoon4
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Re: First attempt at PF-tek: no mycelium after 2 weeks [Re: jee3apha] * 1
    #27868323 - 07/18/22 10:16 PM (2 years, 5 months ago)

Your cake moisture and consistency looks spot on this time.

Yeah, you can reuse a syringe. I usually use a 10cc syringe on 5 separate occasions. You can also refill a syringe a bunch of times if u make your own spore print. One print can make hundreds of syringes refills.

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Offlinejee3apha
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Re: First attempt at PF-tek: no mycelium after 2 weeks [Re: jee3apha]
    #28032147 - 11/04/22 04:09 AM (2 years, 2 months ago)

For posterity, it seems the issue was a bad spore syringe after all. Thank you guys for looking into this.

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