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Offlinehihihi1717
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Re: Accidentally inoc'd 3:1:1 pf jars (12 half pints) [Re: Giggle_Grower]
    #18988787 - 10/16/13 10:37 PM (10 years, 4 months ago)

UPDATE (10/17/13)

The growth I mentioned above is now large enough to get a picture of, so I'll post that here. If anyone can confirm that this is mycelium and not cobweb, that would be fantastic.



It looks rhysomorphic to me, but I'm an amateur.


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Offlinesilverstem
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Re: Accidentally inoc'd 3:1:1 pf jars (12 half pints) [Re: hihihi1717]
    #18988805 - 10/16/13 10:40 PM (10 years, 4 months ago)

i told u to ride it out and u did... sooo goood job thats myc! keep it up buddy.


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Offlinehihihi1717
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Re: Accidentally inoc'd 3:1:1 pf jars (12 half pints) [Re: silverstem]
    #18988831 - 10/16/13 10:45 PM (10 years, 4 months ago)

Awesome! The only thing that's a bit of a bummer is that this is the only jar showing growth after 5 days, but I do understand that it can take weeks depending on the variables of this experiment. Strain, temp, substrate, etc.

This is also the only part of the jar showing growth at the inoculation point, if it's the only point of origin I'll get, how long can it take for a 1/2 pint jar to colonize like this (again, I know it's different for everyone but a ballpark maximum would be appreciated :laugh:)?


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OfflineSockadin
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Re: Accidentally inoc'd 3:1:1 pf jars (12 half pints) [Re: hihihi1717]
    #18988834 - 10/16/13 10:46 PM (10 years, 4 months ago)

Awesome Vid.  I love it.


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Offlinesilverstem
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Re: Accidentally inoc'd 3:1:1 pf jars (12 half pints) [Re: Sockadin]
    #18988851 - 10/16/13 10:52 PM (10 years, 4 months ago)

its normal when using ms to not see growth within the first week... some get lucky and see growth in 3 days. for other it may take a week... now idk how long its gonna take because u are using less brf then the normal recipe calls for so less nutrients may cause it to stall or to spread less quickly then normal.


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Offlinehihihi1717
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Re: Accidentally inoc'd 3:1:1 pf jars (12 half pints) [Re: silverstem]
    #18989338 - 10/17/13 01:26 AM (10 years, 4 months ago)

Hmm, "ms"? What's "ms"?


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InvisiblePestile

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Re: Accidentally inoc'd 3:1:1 pf jars (12 half pints) [Re: hihihi1717]
    #18989405 - 10/17/13 01:59 AM (10 years, 4 months ago)

Multi spore.


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Offlinehihihi1717
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Re: Accidentally inoc'd 3:1:1 pf jars (12 half pints) [Re: Pestile]
    #18989412 - 10/17/13 02:02 AM (10 years, 4 months ago)

When wouldn't there be multiple spores? We're talking about sexual reproduction, right? Not asexual?


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InvisiblePestile

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Re: Accidentally inoc'd 3:1:1 pf jars (12 half pints) [Re: hihihi1717]
    #18989429 - 10/17/13 02:10 AM (10 years, 4 months ago)

When we're talking about an isolate. Then it's living mycelium which has gone through transfer after transfer until it is a mono-culture. No sexual reproduction.


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Offlinehihihi1717
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Re: Accidentally inoc'd 3:1:1 pf jars (12 half pints) [Re: Pestile]
    #18989476 - 10/17/13 03:07 AM (10 years, 4 months ago)

So, in theory, one spore could inoculate an entire jar? However unlikely?


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OfflineGiggle_Grower
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Re: Accidentally inoc'd 3:1:1 pf jars (12 half pints) [Re: hihihi1717]
    #18989569 - 10/17/13 05:06 AM (10 years, 4 months ago)

No. You need at least two spores for a culture. And isolate is when you already have a culture with bunches of genetics, then you selcect the best growth, let it grow, select again and repeat until you have a single set of genetics. A monoculture.


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InvisiblebodhisattaMDiscordReddit
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Re: Accidentally inoc'd 3:1:1 pf jars (12 half pints) [Re: hihihi1717]
    #18990072 - 10/17/13 09:37 AM (10 years, 4 months ago)

Quote:

hihihi1717 said:
When wouldn't there be multiple spores? We're talking about sexual reproduction, right? Not asexual?




Quote:

hihihi1717 said:
So, in theory, one spore could inoculate an entire jar? However unlikely?




If you look at a picture of MS syringe (a very small drop) on an agar plate you'll see "sectors" of growth. If you take a sliver from one sector and put that on a new dish you eliminated 90+% of the genetic diversity of that one drop of MS solution(think about all the diversity in 1/4cc!!) now that new wedge will grow out and probably still show a bunch of sectors. Take one sector again and put that on a new dish. At this point if you're lucky you'll have an isolate or you'll need to keep going to see no more sectors. Alternatively you could take clean tissue from the inside of a fruit and put that to agar and end up with only the 1 mushrooms genetics. You can keep all the genetics of that shroom or isolate from that too. Then you can use those isolates to inoculate a whole jar without spores/genetic diversity.

Watch this video if you're interested it's not long.

If your REALLY interested lectures 2,3,and 12 have some info too
Mycology lectures



Edited by Trusted cuItivator (10/17/13 09:57 AM)


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Offlinehihihi1717
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Re: Accidentally inoc'd 3:1:1 pf jars (12 half pints) [Re: bodhisatta]
    #18991786 - 10/17/13 04:38 PM (10 years, 4 months ago)

That's some pretty neat stuff!

I'll have to try agar after I've had some success with the pf tek and maybe another.

Also, as an update, jar 2 is growing rather nicely (twice the size since I found its growth 2 days ago), and jars 1 and 6 now have signs of life. Each jar has mycelial growth at one of the inoculation points. I suspect 3:1:1 ratios aren't going to ruin substrate unless I lose these jars to stalling due to a lack of nutrients. I'll need to work on raising the temperature of my room, though. My room probably sits at 69-70 degrees when I'm not in it. Thankfully a lot of the time me and my friends are LANing so that raises the temperature quite a bit. Time will tell!


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OfflineGiggle_Grower
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Re: Accidentally inoc'd 3:1:1 pf jars (12 half pints) [Re: hihihi1717]
    #18992801 - 10/17/13 08:05 PM (10 years, 4 months ago)

Your temps are just fine. Higher temps increase chance of contams.


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Offlinehihihi1717
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Re: Accidentally inoc'd 3:1:1 pf jars (12 half pints) [Re: Giggle_Grower]
    #18995413 - 10/18/13 12:32 PM (10 years, 4 months ago)

Okay, so quick update with a follow up question.

Jar #2 now has growth at two inoculation points! :grin:

However, in the first inoculation point, it has begun to colonize into the dry verm layer as well as the regular brf substrate. The point of origin for the mycelium appears to be only a few centimeters at most from the dry verm layer, so I suppose this isn't an outlandish thing to happen. Will it change anything, though? Contamination/nutrition wise? And will I need to alter my methods for the dunk and roll/fruiting chamber set up in any way?

Super stoked about the growth at a second inoculation point, though!

I can't thank you all enough for your help in this, by the way. This is quite an experience already, and I haven't even gotten that far!


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OfflineMush4Brains
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Re: Accidentally inoc'd 3:1:1 pf jars (12 half pints) [Re: Giggle_Grower]
    #18995432 - 10/18/13 12:37 PM (10 years, 4 months ago)

Quote:

Giggle_Grower said:
No. You need at least two spores for a culture. And isolate is when you already have a culture with bunches of genetics, then you selcect the best growth, let it grow, select again and repeat until you have a single set of genetics. A monoculture.




Not really, you just won't get fruiting with monokarytoic mycelium.  Theoretically, it could cover a whole jar.


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InvisiblePestile

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Re: Accidentally inoc'd 3:1:1 pf jars (12 half pints) [Re: hihihi1717]
    #18995437 - 10/18/13 12:39 PM (10 years, 4 months ago)

The dry verm layer will be riddled with mycelium when you birth it. It's normal.


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Offlinehihihi1717
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Re: Accidentally inoc'd 3:1:1 pf jars (12 half pints) [Re: Pestile]
    #18995502 - 10/18/13 12:54 PM (10 years, 4 months ago)

Excellent!

I'm also curious. If my other jars don't take, is it possible to take a tiny piece of mycelium from fully colonized jars prior to birthing to reinoculate the jars that don't grow? As a sort of jump-start? Or is that just a waste of cake?


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InvisiblePestile

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Re: Accidentally inoc'd 3:1:1 pf jars (12 half pints) [Re: hihihi1717]
    #18995531 - 10/18/13 01:01 PM (10 years, 4 months ago)

No, that would not work.


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Offlinehihihi1717
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Re: Accidentally inoc'd 3:1:1 pf jars (12 half pints) [Re: Pestile]
    #19075280 - 11/02/13 01:59 PM (10 years, 3 months ago)

Update time!

5/12 jars have begun colonization! Jars 1, 2 and 6 are doing the best, with 1, 3 and 2 inoculation points showing growth, respectively. Jar 1 is doing well with 1 inoculation point, as it is growing very fast and has covered probably 1/3 of the jar. Jars 9 and 12 are also showing growth, albeit delayed (only just showed growth signs a week ago), at 1 inoculation point, each.

I'll post some pictures below. I'm worried about one of the jars, though, which is jar #2. It has the 3 inoculation points, but it appears to have two or three metabolites present, and I'm just worried that they might be contaminated. Pics below...

Jar #1



Jar #2





Jar #6





Jar #9



Jar #12



So, how are they looking to you guys? Thanks again for all the help!


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