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OfflineShr00merN00ber
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18 Petris, Frist Timer some insight needed!
    #26411003 - 01/01/20 01:54 PM (4 years, 28 days ago)

Hey guys, happy NY to all! So here we go, my first time doing this properly. I've got 18 petris going. I took tissue samples from the best shrooms growing out of a growbox. There were 3 initial samples, I took a tissue sample from a big mushroom growing out of a cluster hereafter called 'STR', and two different tissue samples from the biggest mushroom hereafter called 'CL1' and 'CL2'. All these dishes are T1 after mycelium grew out from the initial samples where I tried to choose the best sectors.

So I have 2 parts to this post.

1) My pressure cooker only fits 2 quart-sized jars... I'm following this tek, is it up to date? https://www.shroomery.org/forums/showflat.php/Number/20048765 What is the preferred option, prep enough grains for 2 jars at a time so there is no waiting after grains are prepped and pressure cooked? Basically, can I load prepped grains in jars and wait a couple of days till the pressure cooker gets through 2 jars at a time?

2) I would like to know your opinion on the plates. I the dishes numbered written on the dish itself. Is the dish G2G worthy? Single grain jar worthy? Further agar transfer needed? Trash worthy?


STR (4 plates)



CL2 (6 plates)



CL 1 (9 plates <I put two photos of #4 because imo could be the best of the lot, maybe good for G2G?>)



Cheers all, happy New Year again!


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OfflineA.k.aM
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Re: 18 Petris, Frist Timer some insight needed! [Re: Shr00merN00ber]
    #26411011 - 01/01/20 02:03 PM (4 years, 28 days ago)

You should probably clear those lids up then repost.

They look decent from what I can tell.
Just set something warm on top of the dish for a min then take a picture.


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InvisibleKimble
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Re: 18 Petris, Frist Timer some insight needed! [Re: Shr00merN00ber]
    #26411033 - 01/01/20 02:20 PM (4 years, 28 days ago)

I personally would do at least another transfer or two as you are looking for healthy uniform appearance.

I'll mention I found filtering the agar mixture before putting it in the pc can reduce the bits of non homogeneous agar that give the plates a spotty appearance. This helps to give a nice clear view of the culture. Also pouring them a wee bit thinner can also help give a nice clear uniform look.

All that said, they look good. No obvious contams I can see. You're on the right track.

I hope that helps.
👍


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OfflineShr00merN00ber
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Re: 18 Petris, Frist Timer some insight needed! [Re: Kimble]
    #26411751 - 01/02/20 12:59 AM (4 years, 27 days ago)

Thanks for the input. Regarding the grains, what shall I do? My pc only fits 2 jars at a time.. Can I leave jars with prepped grains waiting? I'm thinking to do the following:

1. Start rinse/soak for about 16 hours.
2. Boil 10 mins, strain, dry and load jars.
3. Start pc with 2 jars 90 mins@15psi, the other jars left in sab.
4. Wait another hour? for jars to cool in pc and repeat pc with next 2 jars and so on.

Will I be risking contams if I do say 10 jars? That will be about 10 hours for the last two jars sitting around before being pc.


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OfflineRandalf the Grey
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Re: 18 Petris, Frist Timer some insight needed! [Re: Shr00merN00ber]
    #26411785 - 01/02/20 02:04 AM (4 years, 27 days ago)

A 10 hour wait won't destroy your grain. Best case would be the seal\wrap jars and keep them in refrigerator until you are ready to of them.


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Randalf the Grey
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Randalf's Book of Genesis: In The Beginning - Teks, ratios, recipes and log entries
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OfflineShr00merN00ber
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Re: 18 Petris, Frist Timer some insight needed! [Re: Randalf the Grey]
    #26411790 - 01/02/20 02:19 AM (4 years, 27 days ago)

Thanks for the reply. So after drying grains, load them in jars, seal and leave in fridge till their turn in the pc? Or do you mean after pc store in fridge till they get inoculated later on that day?


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OfflineRandalf the Grey
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Re: 18 Petris, Frist Timer some insight needed! [Re: Shr00merN00ber]
    #26411794 - 01/02/20 02:27 AM (4 years, 27 days ago)

In fridge until PC. After PC, sit them in SAB while they cool if they are still hot you you take them out. Keep foil on top until ready to noc up. Once PC'd, there shouldn't be any organisms left in the grain to cause spoilage\rot. They will likely be fine just sitting on a counter but I always try to play it safe. Putting them in SAB just limits the amount of contams the outside of jar and filter will be exposed to while it cools.
This is personal experience\prefernce talking so please don't take my word as gospel.


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Randalf the Grey
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OfflineShr00merN00ber
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Re: 18 Petris, Frist Timer some insight needed! [Re: Randalf the Grey]
    #26411799 - 01/02/20 02:35 AM (4 years, 27 days ago)

Awesome, thanks so much. Will be starting tomorrow. Any petris you see that you like for g2g or particularly dislike? Also, what do you think is best practice, 2 jars per petri dish or 1?


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OfflineRandalf the Grey
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Re: 18 Petris, Frist Timer some insight needed! [Re: Shr00merN00ber]
    #26411832 - 01/02/20 03:13 AM (4 years, 27 days ago)

Depends on jar size. I only use pints or half pints for G2G (easier to work with in SAB). I put 2-5 wedges per jar.
I can't see well enough to point out sectors to cut but can see that they have some uneven growth. I would do at least 1 more transfer before starting a master grain jar. Get a full plate that shows even across the whole thing and you can get 5-10 jars out of it. If youre in a hurry, just do one jar from the best looking edge of one plate while you do more transfers to be safe.


--------------------
Randalf the Grey
Lost in the right direction
A wizard is never late..........but I can try
Randalf's Book of Genesis: In The Beginning - Teks, ratios, recipes and log entries
LAGM2O2O Grow Log
Genesis II: Electric Boogaloo - The Restart


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OfflineShr00merN00ber
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Re: 18 Petris, Frist Timer some insight needed! [Re: Randalf the Grey]
    #26411839 - 01/02/20 03:24 AM (4 years, 27 days ago)

Awesome, thanks so much Randalf. To be honest, I'm surprised you guys can't see the petris clearly, there are a few that the moisture beads can be considered an issue in determining growth but I think most are pretty clear no? In any case, thanks again dude!


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OfflineRandalf the Grey
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Re: 18 Petris, Frist Timer some insight needed! [Re: Shr00merN00ber]
    #26411841 - 01/02/20 03:27 AM (4 years, 27 days ago)

Happy to help. :smile:
I think it's more of a focal length issue than condensation. Camera seems to have focused on plates\agar instead of myc. Just not enough detail to give accurate answers.

Also, my eyes suck!


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Randalf the Grey
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Randalf's Book of Genesis: In The Beginning - Teks, ratios, recipes and log entries
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Edited by Randalf the Grey (01/02/20 03:27 AM)


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InvisibleSpunkyMonkey88
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Re: 18 Petris, Frist Timer some insight needed! [Re: Randalf the Grey]
    #26411918 - 01/02/20 05:59 AM (4 years, 27 days ago)

Quote:

Randalf the Grey said:
Depends on jar size. I only use pints or half pints for G2G (easier to work with in SAB). I put 2-5 wedges per jar.
I can't see well enough to point out sectors to cut but can see that they have some uneven growth. I would do at least 1 more transfer before starting a master grain jar. Get a full plate that shows even across the whole thing and you can get 5-10 jars out of it. If youre in a hurry, just do one jar from the best looking edge of one plate while you do more transfers to be safe.




What do you mean you put 2-5 wedges per jar?

You dont just cut a plate into 1/4ths or halves and drop a section in?


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OfflineRandalf the Grey
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Re: 18 Petris, Frist Timer some insight needed! [Re: SpunkyMonkey88]
    #26412266 - 01/02/20 10:30 AM (4 years, 27 days ago)

I suppose that is an option, but not how I do it.
Each piece of culture that you put in a jar is another point of inoculation. More wedges means more points which means faster colonization. if you have one giant Chunk in a jar and you shake it you're not going to touch as many pieces in the grain as its you have five smaller pieces and shake it.
there is another method of using very soft agar. Soft enough that when it is shaken, it almost reliquify eyes and coats all the grain. All of these are options.
It also depends a bit on the size of your Petri dishes. You will be able to inoculate more grain jars with a 100 mm petri dish than a 60 mm


--------------------
Randalf the Grey
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Randalf's Book of Genesis: In The Beginning - Teks, ratios, recipes and log entries
LAGM2O2O Grow Log
Genesis II: Electric Boogaloo - The Restart


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OfflineFonzee
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Re: 18 Petris, Frist Timer some insight needed! [Re: Randalf the Grey]
    #26412303 - 01/02/20 10:52 AM (4 years, 27 days ago)

I'm no pro, but I think you want to take much smaller bits for cloning. Your cultures don't seem too uniform, and thus might require a bit of further isolation before going into mass production with them.
The growth seems healthy to me.


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OfflineShr00merN00ber
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Re: 18 Petris, Frist Timer some insight needed! [Re: Fonzee]
    #26412329 - 01/02/20 11:06 AM (4 years, 27 days ago)

Hey guys, thanks for the input again. The agar sample taken from the initial dish with live mushroom tissue was pretty small, about 1/2 cm.

The areas in the dish which seem like an empty circle (eg plate 3 from CL1 between 23-2 o clock) will eventually be filled by stronger mycelium?


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Offlineverum subsequentis
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Re: 18 Petris, Frist Timer some insight needed! [Re: Shr00merN00ber]
    #26412484 - 01/02/20 12:53 PM (4 years, 27 days ago)

I don't see any obvious problems but it's impossible to tell if they are clean because the pics aren't good enough.

To clear up a comment above, someone said "they aren't uniform enough". This is a bit of a misnomer. Plates don't have to be uniform to be clean. It's good to go for uniform organized cultures but not always necessary. Cultures with genetics variety can grow all kinds of "not uniform" and still be clean.

You asked if any of the cultures are good enough for G2G. This is also a misnomer. G2G means grain to grain.... I think you mean "are any of these plates good enough for A2G (agar 2 grain)". The answer to that is probably but no guarantees.

You can load jars and leave them sitting at room temp for ten hours without any issues. I wouldn't bother wrapping them or fridgeing them. I routinely make up more grain than my pc can handle and do it in batches. In fact, as soon as i get off the toilet I'm gonna go start straining and bagging grain. It'll be enough to fully load my PC twice. One load goes in and the other just sits on the counter until the pc is free.


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Offlinefootpath
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Re: 18 Petris, Frist Timer some insight needed! [Re: verum subsequentis]
    #26412556 - 01/02/20 01:40 PM (4 years, 27 days ago)

As far as uniformity goes, isn't it still a good indication if there is obvious obstruction or restriction of growth? Case to case, of course.. for instance, in this case I would say that the growth on all of those plates is well within a uniform growth - maybe not perfectly circular, but uniform and unimpeded nonetheless.

But if you have an almost perfect circle of growth with a very obvious divot in one portion of the growth - would you not say that would likely indicate contamination?

I always try to imagine plates in time-lapse as best as I can.
I feel like it's kind of aspect of learning to see, if you can take any given snapshot and predict the previous growth process. I feel like those plates lacking in that certain uniformity are ones that you can fairly easily draw back to an earlier date and subsequently diagnose.
Maybe I'm crazy though.


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Offlineverum subsequentis
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Re: 18 Petris, Frist Timer some insight needed! [Re: footpath]
    #26412602 - 01/02/20 02:01 PM (4 years, 27 days ago)

You're not crazy. That all makes sense.

There are just too many possibilities and variables to say yes or no. In general, it's a good idea for noobs to look for uniformity but in reality it's an absurd over simplification. Sometimes that's a lot easier than diagnosing every particular noob plate ever uploaded with shitloads of condensation and bad focal points. But, alas, I can't bring myself to give the simple answer.

Plates can be wildly "un uniform" and still clean. It all depends on what's going on in the particular plate. Very nice and clear pictures are needed for one to help online. And even then, the cultivator simply has to learn to see what's going on.

But, yes, i do agree with everything you stated.


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OfflineA.k.aM
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Re: 18 Petris, Frist Timer some insight needed! [Re: verum subsequentis]
    #26412623 - 01/02/20 02:12 PM (4 years, 27 days ago)

I always thought uniformity was a sign that it was clean, but mostly that enough strains were removed that if you grow it out it’ll be super consistent.

It’s insane how much there is to learn once you get into agar


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Edited by A.k.a (01/02/20 02:13 PM)


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OfflineShr00merN00ber
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Re: 18 Petris, Frist Timer some insight needed! [Re: A.k.a]
    #26412829 - 01/02/20 03:53 PM (4 years, 27 days ago)

Thanks for the clarification and knowledge verum. I'll keep you guys posted and upload some better pics. Going for the first plate to 2 quarts of rye in my SAB to see whats up. I think I'm going to divide the mycelium into 6 parts, 3 per jar. Most likely going for CL2 #4. Just smoked a fat one too, really excited!


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Offlineverum subsequentis
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Re: 18 Petris, Frist Timer some insight needed! [Re: Shr00merN00ber]
    #26413118 - 01/02/20 07:14 PM (4 years, 27 days ago)

:fuckyeah:
you got this


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Offlineverum subsequentis
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Re: 18 Petris, Frist Timer some insight needed! [Re: verum subsequentis]
    #26413216 - 01/02/20 08:25 PM (4 years, 27 days ago)



Just to show you. These were bagged up a couple hours ago and will remain there overnight and well into tomorrow. Unsealed and not in the fridge. I'll load them up and cook them whenever I get around to it tomorrow. They'll be fine


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OfflineRandalf the Grey
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Re: 18 Petris, Frist Timer some insight needed! [Re: verum subsequentis]
    #26413220 - 01/02/20 08:27 PM (4 years, 27 days ago)

That is a beautiful sight verum!  Very pleasing to look at.

Definitely listen to verum over ANYTHING I said. :smile: And rock on man!


--------------------
Randalf the Grey
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Randalf's Book of Genesis: In The Beginning - Teks, ratios, recipes and log entries
LAGM2O2O Grow Log
Genesis II: Electric Boogaloo - The Restart


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OfflineShr00merN00ber
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Re: 18 Petris, Frist Timer some insight needed! [Re: Randalf the Grey]
    #26413659 - 01/03/20 05:13 AM (4 years, 26 days ago)

Awesome looking bags verum, a real pro. Is that wbs? Here is a pic of one of my jars today, all grains were dried aprox an hour, they were dry to the touch and no moisture on paper towel when left on for 20 seconds. How's it looking?



What I found pretty interestingly hard about the process was the fact that the agar wedges love to stick to the glass. Pretty annoying as I would have liked them to be in the center, covered by some grain. On one jar I had to tilt the grains onto the side to have contact with the two wedges that were stuck on the glass.  :cockafuck:


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Offlineverum subsequentis
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Re: 18 Petris, Frist Timer some insight needed! [Re: Shr00merN00ber]
    #26413692 - 01/03/20 06:02 AM (4 years, 26 days ago)

Your grain looks sexy as midget tits.

Agar can stick to the glass, for sure. It's a super duper boner buster until you figure out how to roll. When i put agar to grain jars, I do as many as i've got without bothering about the placement of the agar. Once all jars are impregnated and covered (re lidded), i do a gently cart wheel with each jar. I say gentle because if the agar hits the wall with much force, it gets emotionally involved and doesn't like to let go.

A few tries and you'll figure it out. Gentle cart wheel and when i'm 3/4 of the way through the flip, i give her a little wiggle as i finish the rotation.

If you don't have a smasher yet, you'll want to get that figured out. I( smash on a big ass wad of cardboard but people use tires or basketballs or..... whatever. If the wedge sticks to the wall, several powerful slams down on the smasher will almost always slurp the agar down the wall and get it under the grain.


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OfflineShr00merN00ber
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Re: 18 Petris, Frist Timer some insight needed! [Re: verum subsequentis]
    #26414965 - 01/03/20 06:58 PM (4 years, 26 days ago)

Damn verum, that roll tek really helped out! I got the slow rhythm of the roll at the end after a couple of jars. Wedges generally all covered by rye. I guess it's in the simple things that experience really matters too. Cheers verum!


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Offlineverum subsequentis
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Re: 18 Petris, Frist Timer some insight needed! [Re: Shr00merN00ber]
    #26415044 - 01/03/20 07:38 PM (4 years, 26 days ago)

Cheers. Yeah, It's all the little things you pick up after years of doing thousands of jars..... OR when someone kind comes along and drops a little cheat on you, saving you years and thousands of jars.

:douchewink:

Glad it made sense for ya.


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OfflineA.k.aM
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Re: 18 Petris, Frist Timer some insight needed! [Re: verum subsequentis]
    #26415144 - 01/03/20 08:39 PM (4 years, 26 days ago)

Gonna try this roll tomorrow. The first time I dropped agar half of it got fused to the glass above the seeds and I ended up just leaving the jar on its side.

I’ve only done two and the second one I dropped them in the middle then tilted the jar just a tiny bit and slapped the side so the seeds jump and it worked ok but still not great.


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Offlineverum subsequentis
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Re: 18 Petris, Frist Timer some insight needed! [Re: A.k.a]
    #26415158 - 01/03/20 08:46 PM (4 years, 26 days ago)

If you roll properly and it still sticks, I'd say you are probably making your agar a little soft. Slightly increasing the agar ratio could help. But probably, you just need a little practice rolling.


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Invisiblemorty422
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Re: 18 Petris, Frist Timer some insight needed! [Re: verum subsequentis]
    #26415185 - 01/03/20 09:09 PM (4 years, 26 days ago)

If this is your absolute first time grow - I'd like to extend a big high five.

You have LOTS of plates, when you start thinking about taking samples off of those plates you start to get pretty intense with things.

I'd say once you have a nice clean cut of each plate you want to keep, go two to agar and one plate to grain. (3 plates of each)

Comeback graced me with that idea and it's kept my grows flowing well. You always have something to do, always have something to put to grain, you never have to really be idle if you don't want to be.

Your plate pictures are very hard to see - I would also say the pieces of tissue grabbed are sort of big - but nothing too out of the ordinary. I'd try to go a little bit smaller though if I were you.

But I'm looking into seeing the rest of this one! I've got a few tubs I've had going for a few days, getting antsy and feeling the pressure just like many do!


:cheers: and good luck!


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OfflineShr00merN00ber
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Re: 18 Petris, Frist Timer some insight needed! [Re: morty422]
    #26415618 - 01/04/20 07:50 AM (4 years, 25 days ago)

Damn, already used most of the good plates. That's an excellent strategy to be adopted morty, wish I had seen it sooner.. Thank you guys for these tips, they are really efficient and practical!


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OfflineShr00merN00ber
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Re: 18 Petris, Frist Timer some insight needed! [Re: Shr00merN00ber]
    #26420580 - 01/07/20 08:49 AM (4 years, 22 days ago)

Hey guys, small update. Here are two of my jars. One of them is quart size, the other is 1/2 quart. My pc fits 2 quart size jars or 8 1/2 quart jars, so opted to get more of those. I'm just worried about moisture content. I don't really see any condensation in most jars, is that a problem? I can't remember where I read it but it was from a TC and they said there should be some visible moisture in the jar, especially when myc is colonizing.  :shrug:  So I opted for 1/2 an agar dish split into 3 per 1/2 quart jar.



To be honest, they looker drier than that irl. 

Cheers!


Edited by Shr00merN00ber (01/07/20 09:00 AM)


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InvisibleKimble
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Re: 18 Petris, Frist Timer some insight needed! [Re: Shr00merN00ber]
    #26420698 - 01/07/20 10:10 AM (4 years, 22 days ago)

I'd be
Quote:

Shr00merN00ber said:
Awesome looking bags verum, a real pro. Is that wbs? Here is a pic of one of my jars today, all grains were dried aprox an hour, they were dry to the touch and no moisture on paper towel when left on for 20 seconds. How's it looking?



What I found pretty interestingly hard about the process was the fact that the agar wedges love to stick to the glass. Pretty annoying as I would have liked them to be in the center, covered by some grain. On one jar I had to tilt the grains onto the side to have contact with the two wedges that were stuck on the glass.  :cockafuck:




That jar imho looks perfect. 👌
Personally that is exactly how i like them to look.


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OfflineA.k.aM
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Re: 18 Petris, Frist Timer some insight needed! [Re: Kimble]
    #26420828 - 01/07/20 11:23 AM (4 years, 22 days ago)

Idk oats but looks like the myc loves it.

Good job man.

Condensation in the jar has a lot to do with how warm the room is also.


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Offlinesporecap
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Re: 18 Petris, Frist Timer some insight needed! [Re: A.k.a]
    #26420966 - 01/07/20 12:27 PM (4 years, 22 days ago)

I'll just leave this here because it's so awesome
Quote:

Mr. Alien said:
A lot of people swing the jar or wiggle the jar to bury the wedge but i just:



the end.




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OfflineA.k.aM
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Re: 18 Petris, Frist Timer some insight needed! [Re: sporecap]
    #26421036 - 01/07/20 12:58 PM (4 years, 22 days ago)

I saw that video a while back and remembered it when I started a2g and had problems with wedges sticking to glass.

It works great although I like to hit the side of the jar where the neck starts, seems to be quicker and move it more into the middle of the jar instead of only down.


--------------------
LAGM2020


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