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OfflineMycoSmith
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Registered: 10/29/20
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Stalled Jars! Last 5-10% HELP (New Pics Today)
    #27017705 - 11/02/20 03:00 PM (3 years, 3 months ago)

I have some grain spawn colonizing in quart jars. Up until this point I feel they have been colonizing well and I haven't seen any contams even on the petri dishes they came from. But now I feel as if I've stalled out. My jars definitely don't seem like they are colonizing the last 5-10% all to well. Anything I should be looking for or any ideas as to why?



Edited by MycoSmith (11/05/20 06:34 PM)


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Offlinefraki58
Mycology Learner


Registered: 11/20/19
Posts: 246
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Re: Stalled Jars! Last 5-10% HELP [Re: MycoSmith]
    #27017709 - 11/02/20 03:03 PM (3 years, 3 months ago)

For how long they been like that? Maybe you have some bacteria on those grains and that's why the myc can't colonize it


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If any of the said above is incorrect, please correct me! Still learning here and constructive criticism is the best :smile:


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OfflineMycoSmith
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Re: Stalled Jars! Last 5-10% HELP [Re: fraki58]
    #27017716 - 11/02/20 03:10 PM (3 years, 3 months ago)

I've really only started keeping an eye on them for a few days now. They have had solid growth up until 4-5 days ago. I Sterilized for 2.5 hrs at 15 psi and have 2 layers of micropore tape for gas exchange. They were started from an agar wedge at the end of September. Average daily incubation temp was about 67 degrees.


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Offlinefraki58
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Registered: 11/20/19
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Re: Stalled Jars! Last 5-10% HELP [Re: MycoSmith]
    #27017722 - 11/02/20 03:14 PM (3 years, 3 months ago)

You can shake them and see how well they recover, if they grow again just fine you can use them to bulk! When the myc is healthy it should take ~2 days to fully recover and cover all the grains with myc again


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If any of the said above is incorrect, please correct me! Still learning here and constructive criticism is the best :smile:


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OfflineMycoSmith
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Re: Stalled Jars! Last 5-10% HELP [Re: fraki58]
    #27017727 - 11/02/20 03:19 PM (3 years, 3 months ago)

Yea I'll give that a go, thanks! What is your thoughts of using the healthy mycelium for G2G transferring. If I Sterilized a silver spoon and scooped the visibly healthy grains off the top of one jar into more jars.


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Offlinefraki58
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Registered: 11/20/19
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Re: Stalled Jars! Last 5-10% HELP [Re: MycoSmith]
    #27017731 - 11/02/20 03:22 PM (3 years, 3 months ago)

I if i have questionable grains i would not do G2G, like this jars in particular. You could start more jars and if you are sure sure they are healthy then you can G2G, else you will be growing the contams in the original jars on your subsequents jars. For transfering the grains there are some videos on how to do it, i'll look for them and come back
Edit: Can't find them right now, but you should be good shaking them prior the G2G to loosen the grains, then you can simply open the recevieng jars and just letting the grains fall in the jar in a smooth move and your jars are ready.


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If any of the said above is incorrect, please correct me! Still learning here and constructive criticism is the best :smile:


Edited by fraki58 (11/02/20 03:28 PM)


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OfflineMycoSmith
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Re: Stalled Jars! Last 5-10% HELP [Re: fraki58]
    #27017743 - 11/02/20 03:34 PM (3 years, 3 months ago)

Thanks for the advice. I will shake a couple up and watch what those and the others do. Fingers crossed I don't have to go back to the plates.


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InvisibleIdiotCircusBoy
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Registered: 09/22/20
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Re: Stalled Jars! Last 5-10% HELP [Re: MycoSmith]
    #27017747 - 11/02/20 03:36 PM (3 years, 3 months ago)

I'm new at this but have you tried using larger text??

Hmm


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Just call me Idiot


"People hasten to judge in order not to be judged themselves."


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OfflineMycoSmith
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Re: Stalled Jars! Last 5-10% HELP [Re: IdiotCircusBoy] * 1
    #27017752 - 11/02/20 03:40 PM (3 years, 3 months ago)

Are you asking me how or asking me to?


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InvisibleIdiotCircusBoy
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Re: Stalled Jars! Last 5-10% HELP [Re: MycoSmith]
    #27017760 - 11/02/20 03:46 PM (3 years, 3 months ago)

Just f*ing around.  Never seen anyone use such large text before.  Interested to see how the grains work out.  Am in similar situation.  Good luck brother!!


--------------------
Just call me Idiot


"People hasten to judge in order not to be judged themselves."


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OfflineMycoSmith
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Re: Stalled Jars! Last 5-10% HELP [Re: IdiotCircusBoy]
    #27017764 - 11/02/20 03:49 PM (3 years, 3 months ago)

Good luck to you as well. What kind of grain are you using?


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OfflineMycoSmith
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Re: Stalled Jars! Last 5-10% HELP [Re: fraki58]
    #27017780 - 11/02/20 04:00 PM (3 years, 3 months ago)

If I were to shake the jars and the uncolonized grain IS contaminated what if the contaminated grains are no longer visible (like they end up in the middle or something) and the rest of the jar recovers and looks fine...is this possible? Or would shaking spread the contamination enough for it to become prevalent and obviously the culprit.


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Offlinefraki58
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Registered: 11/20/19
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Re: Stalled Jars! Last 5-10% HELP [Re: MycoSmith]
    #27017790 - 11/02/20 04:10 PM (3 years, 3 months ago)

You will see once it recovers, if there is a little bacteria maybe you wont even see it in the jar, you can still use bacterial spawn for bulk, people here have used it and it works, it will impact on the final yield, but, if you want some fruits you prolly can get them anyways!


--------------------
If any of the said above is incorrect, please correct me! Still learning here and constructive criticism is the best :smile:


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OfflineMycoSmith
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Re: Stalled Jars! Last 5-10% HELP [Re: fraki58]
    #27017799 - 11/02/20 04:20 PM (3 years, 3 months ago)

Thanks for the new knowledge on bacterial contamination👍 I'm going to shake half of them and let the other half stay as is.


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Offlinedirtybirdx
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Re: Stalled Jars! Last 5-10% HELP [Re: MycoSmith] * 1
    #27017817 - 11/02/20 04:31 PM (3 years, 3 months ago)

Quote:

MycoSmith said:
Thanks for the new knowledge on bacterial contamination👍 I'm going to shake half of them and let the other half stay as is.





You should shake them all.

There will be no benefit to leaving them.

They are either going to:

A) not colonize

or

B) take so long to colonize that the myc gets stressed and bacterial, with enough metabolites to make you want to throw it out.


When you shake, they are either going to:

A) recover with a bit of bacteria, usable.

or

B) recover with a lot of bacteria or contam


—

Those jars are already not clean.  Waiting, shaking, not waiting,,,,nothing will change that.  I’ve had enough of those to know.


--------------------
Need help?




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Offlinefraki58
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Registered: 11/20/19
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Re: Stalled Jars! Last 5-10% HELP [Re: dirtybirdx]
    #27017861 - 11/02/20 04:57 PM (3 years, 3 months ago)

:whathesaid:


--------------------
If any of the said above is incorrect, please correct me! Still learning here and constructive criticism is the best :smile:


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OfflineMycoSmith
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Re: Stalled Jars! Last 5-10% HELP [Re: dirtybirdx]
    #27017871 - 11/02/20 05:02 PM (3 years, 3 months ago)

Quote:

dirtybirdx said:
Quote:

MycoSmith said:
Thanks for the new knowledge on bacterial contamination�� I'm going to shake half of them and let the other half stay as is.





You should shake them all.

There will be no benefit to leaving them.

They are either going to:

A) not colonize

or

B) take so long to colonize that the myc gets stressed and bacterial, with enough metabolites to make you want to throw it out.


When you shake, they are either going to:

A) recover with a bit of bacteria, usable.

or

B) recover with a lot of bacteria or contam


—

Those jars are already not clean.  Waiting, shaking, not waiting,,,,nothing will change that.  I’ve had enough of those to know.




Okay I have shaken them all I hope for a good recovery here is what a few  of em look like






Before




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OfflineMycoSmith
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Registered: 10/29/20
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Re: Stalled Jars! Last 5-10% HELP [Re: MycoSmith]
    #27017908 - 11/02/20 05:25 PM (3 years, 3 months ago)

So if all goes well and these jars produce fruits when spawned to 100% coir can I still clone one of the fruits? or is cloning a fruit not a good idea if a bacterial contamination is suspected


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Offlinetiptrippy
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Registered: 09/09/20
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Re: Stalled Jars! Last 5-10% HELP [Re: MycoSmith]
    #27017939 - 11/02/20 05:41 PM (3 years, 3 months ago)

From my understanding, you can clone a fruit that was produced with bacterial spawn on Agar. Watch the growth and transfer a healthy uncontaminated section on to a new Agar plate a few times and you will end up with a healthy clone plate that you can use to inoculate more grain.


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OfflineMycoSmith
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Re: Stalled Jars! Last 5-10% HELP [Re: tiptrippy]
    #27018014 - 11/02/20 06:05 PM (3 years, 3 months ago)

Okay thank you for this information. I am hoping it is just a combination of multiple genetics and the colder incubation temperatures
(62-68 degrees). I will chime back in with new pics if they recover nicely. Also I purchased an aquarium heater today to make an incubation tub for my next round. I will be making some no pour agar dishes for a b+ syringe I have.

These jars came from 3-4 different petri dishes which none I had contaminations in the whole batch, not just the transfer plates... What are the odds that the slow growth is from the constant sub 70 degrees incubation and not a bacterial contam? If they recover nicely how do I know if they are clean or just "look clean" with a small bacterial contam


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