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Re: Agar culture question [Re: Funnguiy]
    #25829072 -

Impatience is a major thing for noobs in this practice. A plate should grow out fully in way less than a month. If it's really moving slowly then there is probably a bacteria problem but i think you are probably just new and impatient.

Small pieces of agar will  colonize a jar without a problem. If spores can do it then obviously the tiniest piece of myc can too. You don't want to put uncolonized agar in the jar but it's not a problem if you do. There is just no point and It's possible that a hidden contam is in that agar. So, why risk it if the agar has no myc to contribute to growth?

I'd advise posting a picture of the plates you are planning on using. We can tell you a lot more about what is going on if we can see.

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Re: Agar culture question [Re: verum subsequentis]
    #25829089 -

P.S. Agar boilovers can be prevented by only half filling your containers.

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Re: Agar culture question [Re: teladi]
    #25829121 -

i'd say three quarters. I go about 3/4 every time. I suppose it depends on the container though. i use a captain morgan handle with 1000cc agar in it.

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Re: Agar culture question [Re: teladi]
    #25829127 -

teladi said:
P.S. Agar boilovers can be prevented by only half filling your containers.



Boil over can be prevented by using more water in your pc, not letting it get to 15 psi(14  is good) and cooling slowly, no need to only fill half your containers.


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It doesn't matter what i think of you...all that matters is clean spawn
I'm tired do me a favor

I'M A MEMBER FIRST, TC WHEN NEEDED AND MOD WHEN I HAVE TO BE !!!

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Re: Agar culture question [Re: cronicr]
    #25829131 -

Maybe so, but it's common practice in every single lab that I've worked in to half fill media bottles when prepping or reheating agar. Preventing a boil over when reheating a full bottle can be prevented by doing it slowly, and keeping an eye on it. Or, half fill it, set the microwave, and walk away.

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Re: Agar culture question [Re: teladi] * 1
    #25829140 -

Yeah i go likely under 3/4's ish but eventually you just get a feel for shit and end up in an unforgiving cycle of blah:lol:


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It doesn't matter what i think of you...all that matters is clean spawn
I'm tired do me a favor

I'M A MEMBER FIRST, TC WHEN NEEDED AND MOD WHEN I HAVE TO BE !!!

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Re: Agar culture question [Re: cronicr]
    #25829281 -

I like to pour my plates like a week ahead of time or so. Just gives time to see if anything starts growing on that plates that is not mycelium. Unless of course you contaminate it during the inoculation.

But then at least you know your pouring is good but your sterile work ethic on the inoculation needs some work.

I'm sure your transfer will be fine. You have nothing to lose. May or may not have to go back to the syringe and reinoculate a different dish, or the transfer may be wonderful.

Either or, you're not wasting much time.


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:greyalien:



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Re: Agar culture question [Re: Vibe_Enthusiast]
    #25829601 -

If you want speed, do a whole plate for one jar. I usually do the tiger drop (since I use pasty plates) just for simplicity, but the speed is nice too. In your case, since you're using petris, consider using half the plate. That's very doable with a scalpel


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To avoid all evil, to cultivate good, and to cleanse one's mind - this is the teaching of the Buddhas

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Re: Agar culture question [Re: juniperus]
    #25830342 -









These are the plates I’m looking to use as innoculants. The PE seems the most promising. The Pans have been the slackers but their mycelium seems the most robust of the three(rhizomorphic) maybe almost tombestoney. The two pans were taken from the same area from the original plate, so relatively similar genetics I’m hoping. Also, should I avoid taking the center transferred piece into the jar, or not an issue?

I’m looking to knock up two quart jars of rye berries for each strain. So six jars total(which I have waiting).

PS the Chitwan might look a little whispy, but that particular plate had a very thin layer of agar in it (I think it was one of the last to be poured) so I think it’s just the depth that makes it look like that.

Thanks everyone!


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“Time flies. It’s up to you to be the navigator.”

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Re: Agar culture question [Re: Funnguiy]
    #25830360 -









This is a rough sketch of my plan. Blue for one jar, red for the other. I feel it would be best to use one whole culture of pan for each jar because they’re are the smallest.


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“Time flies. It’s up to you to be the navigator.”

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Re: Agar culture question [Re: Funnguiy]
    #25830715 -

I made the transfers, pretty much exactly as outlined above. I think I did a good job, my only concern is that I pre cut the whole culture, stabbed the four pieces like a kabob on my xacto, but they tended to stick to the knife and not fall into the jar. So I swiftly nudged them off with just the tip of my pointer finger (gloved, cleaned with iso beforehand) hopefully that didn’t fuck up all my jars. Each jar has four decent sized pieces.

I shook a couple that had pieces sticking to each other inside, to no avail. I might wait for them to recover and distribute(shake) tomorrow. Or should I do that now?

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Re: Agar culture question [Re: Funnguiy]
    #25830730 -

You fucked up your jars. Hand are not sterile. Learn to use your index finger to tap on your instrument to release the wedge

Edit: you will also have more success if you only transfer one wedge at a time


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CA weed @ all times unless specified
currently this high
pebbles suck
its turtles all the way down

Edited by psilly the kid (02/22/19 06:15 PM)

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Re: Agar culture question [Re: psilly the kid]
    #25830733 -

Well fuck...

If it goes to shit I’m using my syringe for the next jars lol enough of this.

And really? My thinking was get all pieces in there at once would equal less time and amount of times the jar was opened?


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“Time flies. It’s up to you to be the navigator.”

Edited by Funnguiy (02/22/19 06:27 PM)

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Re: Agar culture question [Re: Funnguiy]
    #25830792 -

So I swiftly nudged them off with just the tip of my pointer finger

OOHH, bad move man. It's possible that it'll work out but that's a horrible idea. It's not recommended but it'd be better to use the inside of the rim of the jar to scrape them off. Better yet, don't stab your cultures. I use the scalpel almost like a shovel and scoop up the wedge.

I once tested glove cleanliness by wiping down my glove with iso and then touching the center of a plate. The dimple my finger left bloomed into a rather beautiful bacterial bonanza a couple days later.

But don't give up on agar man. Diving in is the hardest part. It'll be easier and easier each time you do it. I don't even think about it anymore.

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Re: Agar culture question [Re: verum subsequentis]
    #25830835 -

Uhg it was only a split second hopefully they take I really don’t want to start all over :/ how long would you say until I know if it’s contamed?


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“Time flies. It’s up to you to be the navigator.”

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Re: Agar culture question [Re: Funnguiy]
    #25830928 -

If it's bacterial, you may not know until you see poor fruiting performance. Molds can show more quickly, sometimes in the jar, but sometimes they will show up during the first flush, or later. So, no one can say. Start something else going, at least if these fail you will have a head start.

Verum is right though, don't give up! I really becomes second nature pretty quickly.


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To avoid all evil, to cultivate good, and to cleanse one's mind - this is the teaching of the Buddhas

Edited by juniperus (02/22/19 08:21 PM)

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Re: Agar culture question [Re: Funnguiy]
    #25830934 -

could be a while. Post pictures as the jars grow out and we'll let you know what we think. You shouldn't have to start all over. Hopefully you transferred into a new dish before using up the whole plate. If not, you messed up. Always take care to ensure you have the culture saved on agar for future grows.

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Re: Agar culture question [Re: verum subsequentis]
    #25831344 -

I don’t have more plates, so yeah I used it all up lol.  I have six more empty jars but I have to make some sort of lid, I had 6 other nice ones that I ordered on the other jars. Any ideas? I got micropore tape to work with and I think this time I’d just inject with syringe. :shrug: but yeah I’ll post updates.

I won’t completely give up on agar, I’ll use for clones and stuff.

Edit: After doing some reading I’m not really sure what my best course of action is here. If the jarshave that high a chance of being screwed, I might just start over. With agar and everything. I was going to try syringe to grain but I’m reading that’s pretty bad and can fuck up the whole bulk substrate and you may not know until you fruit :sad: definitely don’t want that. I’d much rather wait longer and get better results doing it correctly. But...

Idk friends I just want a long term amount of good quality shrooms, mainly to try Microdosing. Last time I grew (my first time!) I got little over 10ish grams TOTAL from 12 pf jars (my fruiting conditions were trash, I know that now). They were also pretty weak imo, not much visuals or anything (strain was Burma and huatla) but it did give me my favorite so far body high (idk why most dislike this, shit was tight) I have until early May that’s when I’ll be too busy to grow. I really wanted to grow this culture out to fetch a clone fruiting in dubs then figure out which one or two strain I liked the best, and do that in a mono after getting a good specimen from ms. But now, especially if I should start with agar again, I’ll probably only get one grow cycle then, not two://///

Also, I’m guessing you can re use the Petri dishes? Some of them have some scratches in it from my xacto? They are the EZ bio research brand, used once so far. If I can, any tips on how to best clean the plates? I’m going to be an absolute fucking nazi with sterile technique from now on

Edited by Funnguiy (02/23/19 04:02 PM)

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Re: Agar culture question [Re: Funnguiy]
    #25836729 -

Update: after much reading, I decided to make new plates. This time I went with Pastywhytes tek. So I now have 24 (hopefully) clean agar. I PC’d them ~45min 15PSI, but my cooker is small. So I let it depressurization for 10ish minutes then forced the pressure out. I know it’s not ideal, but my cooker could only do batches of 8 at a time. It’s ok tho the last batch has been sitting in it overnight.

Then with some of the new plates, I took cultures from my other transfers on the petris(it grew out on the edges from where I cut the innoculant wedges out). Each strain had two pieces from the same plate go to two of pastwhyte’s plates. Also, just for the hell of it and to get some fresh genetics going, I MS syringe nocced one plate of each strain. THIS time I flame sterilized my xacto and syringe, then in the SAB dropped some solution to cool blade off, and slowly squirted a couple beads onto the blade, and cut a X into the pasty plates with the saturated blade. So, I have three plates of each strain now not including the donor petris. This was done last night so no growth yet, not that I can tell.

I have my sights set on the tiger drop for my next grain jars, I probably won’t do anything with the ones I most likely messed up earlier, I’m too paranoid lol. I liked Pasty’s tek much better, logically imo it makes way more sense to sterilize afterrrr you pour, but that’s just me. Eventually I’ll learn proper pour technique, but for now I need something a little more fool proof. I am curious to see what my contam rates will be for this. Here are some photos, check out that blue!!





The 500ml bottle I had filled with agar had JUST enough for all 24, which was dope and satisfied my slight OCD when it comes to mycology.



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“Time flies. It’s up to you to be the navigator.”

Edited by Funnguiy (02/25/19 12:34 PM)

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Re: Agar culture question [Re: Funnguiy]
    #25836764 -

It can take a while for germination depending on how old the spores are. You can expect anywhere from 3-30+ days.

I love pasty plates. Good luck :thumbup:


--------------------
To avoid all evil, to cultivate good, and to cleanse one's mind - this is the teaching of the Buddhas

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