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TheOffice
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The basics of working with Agar
#27307125 - 05/13/21 02:27 PM (3 years, 8 months ago) |
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Hey People,
I've been working with mushrooms for about 4/5 months now. I started working with agar right away, as pretty much everyone here suggests I can say now my preparing and pouring agar is up to par. So after cloning from my first growkit and taking a sporenprint from the best fruit. I'm now going the sporenprint to harvest route. I've swapped some spores on agar, and took three small transfers from the best looking myc. From those three samples I took 3 pieces (transfers) each. So now I ended up with 8 petridishes (one got ruined). https://files.shroomery.org/files/21-19/093500898-IMG-20210513-WA0008.jpg
So now my question, I read people talking about taking 5,6,7 transfers from a sporenprint till rhizomorphic growth or an isolate appear. Now i have no visible contamination (except for number 8, I know, but that is not coming from the transfer)
In number 8,9,10 and 2 there is still sectoring going on, so I will make transfers from them. But number 3,4,5,6,7 don't have sectoring any more
-So are these already isolates? (after two tansfers from a sporenprint)
3,4,5,6,7 have nice healthy growth, but far from rhizomorphic. So what do I do?
Should I transfer from 3,4,5,6,7 also, till I get rhizomorphic growth or just put it on grain/lc like this
https://files.shroomery.org/files/21-19/093569038-IMG_20210513_215328.jpg
I want to do both agar to grain and agar to lc, to find out what works best for me, so it would save me up a bunch of work, grain and time if you guys have any advice for me.
Thanks in advance and keer up the good work!
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Hindsight
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Re: The basics of working with Agar [Re: TheOffice] 1
#27307155 - 05/13/21 03:05 PM (3 years, 8 months ago) |
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Any reason you are growing out a spore print of the best fruit instead of taking a tissue sample to clone from? That would get you more consistent results than rolling the dice again on spores and new genetics.
Are you trying to develop new strains with specific characteristics or are you just trying to establish a reusable culture that fruits well? If the latter, then there is no point in doing a bunch of transfers and selecting for rhizomorphic growth, or starting with spores each time. If you are starting from ground zero with spores, you knock up some plates, take some transfers from those plates, and assuming the transferred plates are all clean, just grow them all out and see what you get. This gives you the widest range of genetics possible, and gives you the best chance at finding something nice in there that does what you are looking for. From there you take a clone of the fruit from tissue sample, then grow it out on agar and/or LC and that becomes your master and you perpetually use it to start grow after grow.
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TheOffice
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Re: The basics of working with Agar [Re: Hindsight]
#27307188 - 05/13/21 03:30 PM (3 years, 8 months ago) |
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Hindsight thanks for the help full and adequate response.
Since I've done the clone route already once (from a grow kit), i thought it would be good for me to start from sporeprint once, to get some experience.
As you say i am not looking for specific characteristics, but for a fast and good fruiter. So if i understand you correctly, the best way to go about is to just put it all to grain as soon as it is all clean. And take tissue samples from the best fruits?
And then one more question, as this happened with the first cloning process. when i take a tissue sample from a fruit and put it to agar. most of the times it grew out into several different sectors. (i understand a fruit from a MS contains more then one genetic) But how do i put this to LC for example? Normally you would take a piece from the leading edge right? but then you can only can take a piece of one specific genetic. Is that desirable (just take the best looking one?) Or do you want to get all of those genetics (sectors) in the LC, by cutting a square out of the middle? if you understand what i am saying.
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TheOffice
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Re: The basics of working with Agar [Re: TheOffice]
#27307197 - 05/13/21 03:37 PM (3 years, 8 months ago) |
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Like this one, this was a growth from a tissue sample. How or what piece of this do I put to grain or lc?
[url=https://files.shroomery.org/files/21-19/094178463-IMG_20210513_233412.jpg
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TheOffice
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Re: The basics of working with Agar [Re: TheOffice]
#27307201 - 05/13/21 03:38 PM (3 years, 8 months ago) |
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Hindsight
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Re: The basics of working with Agar [Re: TheOffice]
#27307212 - 05/13/21 03:50 PM (3 years, 8 months ago) |
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Quote:
TheOffice said: So if i understand you correctly, the best way to go about is to just put it all to grain as soon as it is all clean. And take tissue samples from the best fruits?
Yup! As long as it's clean, grow out the most genetics as possible. It's like having 25 grows in 1. vs isolating 1 plate down to 25 cultures and growing each of the 25 out.
Quote:
TheOffice said: And then one more question, as this happened with the first cloning process. when i take a tissue sample from a fruit and put it to agar. most of the times it grew out into several different sectors. (i understand a fruit from a MS contains more then one genetic) But how do i put this to LC for example? Normally you would take a piece from the leading edge right? but then you can only can take a piece of one specific genetic. Is that desirable (just take the best looking one?) Or do you want to get all of those genetics (sectors) in the LC, by cutting a square out of the middle? if you understand what i am saying.
Correct - a fruit will usually contain several genetics all working together in harmony, but you know whatever the case may be, that these genetics should produce the same fruit again (when grown from clone).
I hear what you are saying about the transfers to LC from a clone-grown agar plate: If there are say 5 different mycelium cultures in that fruit and you put a tissue sample from the fruit onto an agar plate, they may all run in different directions so if you take a wedge from the edge, you might not be getting all the same genetics in your next grow. I take from the middle for that reason, though I do find that my agar plates from clones tend to look fairly uniform - much more so than early MS transfers. This particular question might be better answered from someone with more cloning experience than me - I am fairly new to growing out clones; I've been focused on many MS grows of many different varieties, and then selecting and saving clones, and am just now going back to clone #1 to grow it out.
Edited by Hindsight (05/13/21 03:52 PM)
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sandman420
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Re: The basics of working with Agar [Re: TheOffice]
#27307214 - 05/13/21 03:52 PM (3 years, 8 months ago) |
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first of all sick handwriting...
that last pic of the tissue sample needs some work before it's ready for grain or LC IMO
Id grab around 3 and 6 o'clock for transfers.
You would be well of to take about 10 transfers from this plate, choose the best of the 10 and throw the rest away while you take the goodester one to LCs or grains.
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TheOffice
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Re: The basics of working with Agar [Re: Hindsight]
#27307609 - 05/13/21 10:58 PM (3 years, 8 months ago) |
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https://files.shroomery.org/files/21-19/094178463-IMG_20210513_233412.jpgQuote:
Hindsight said:
Quote:
TheOffice said: So if i understand you correctly, the best way to go about is to just put it all to grain as soon as it is all clean. And take tissue samples from the best fruits?
Yup! As long as it's clean, grow out the most genetics as possible. It's like having 25 grows in 1. vs isolating 1 plate down to 25 cultures and growing each of the 25 out.

Quote:
TheOffice said: And then one more question, as this happened with the first cloning process. when i take a tissue sample from a fruit and put it to agar. most of the times it grew out into several different sectors. (i understand a fruit from a MS contains more then one genetic) But how do i put this to LC for example? Normally you would take a piece from the leading edge right? but then you can only can take a piece of one specific genetic. Is that desirable (just take the best looking one?) Or do you want to get all of those genetics (sectors) in the LC, by cutting a square out of the middle? if you understand what i am saying.
Correct - a fruit will usually contain several genetics all working together in harmony, but you know whatever the case may be, that these genetics should produce the same fruit again (when grown from clone).
I hear what you are saying about the transfers to LC from a clone-grown agar plate: If there are say 5 different mycelium cultures in that fruit and you put a tissue sample from the fruit onto an agar plate, they may all run in different directions so if you take a wedge from the edge, you might not be getting all the same genetics in your next grow. I take from the middle for that reason, though I do find that my agar plates from clones tend to look fairly uniform - much more so than early MS transfers. This particular question might be better answered from someone with more cloning experience than me - I am fairly new to growing out clones; I've been focused on many MS grows of many different varieties, and then selecting and saving clones, and am just now going back to clone #1 to grow it out.
For someone fairly new in growing out clones, your advice is really clear and sounds as the right thing to do:) 
And as far from growing out clones, is this something i can do 'forever'? I understand I can stretch the use of one clone by multiplying my lc or do grain2grain several times. But isn't there going to be a point that I need to start over from sporenprint again, or can I keep taking clones from clones etc?
I've not yet mastered the act of making some kind of master slant where i can start from over and over again. For now my only starting point is agar, and that doesn't keep cultures viable for very long right?
Any advice on how to keep a good culture going?
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TheOffice
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Re: The basics of working with Agar [Re: sandman420]
#27307612 - 05/13/21 11:05 PM (3 years, 8 months ago) |
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Quote:
sandman420 said: first of all sick handwriting...
that last pic of the tissue sample needs some work before it's ready for grain or LC IMO
Id grab around 3 and 6 o'clock for transfers.
You would be well of to take about 10 transfers from this plate, choose the best of the 10 and throw the rest away while you take the goodester one to LCs or grains.
Thanks for the response sandman420!
Clear answer, and will do just that! But just out of curiosity. If this tissue sample that fromed that good fruit, contains several genetics to form that fruit. Wouldn't making transfers from the 3 and 6 o clock area, select out specific genes, therefore not conserving the right combinations of genes to form this fruit?
Or is it more likely that the genes that form the 'good myc sectors' are responsible for the growing of good fruits?
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verum subsequentis
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Re: The basics of working with Agar [Re: TheOffice]
#27307633 - 05/13/21 11:49 PM (3 years, 8 months ago) |
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Don't worry about it. I asked the same questions growing up and soon learned (with enough growing) that there's a damn good reason nobody has a solid answer to that.... Other than, don't worry about it. Just snag three or four of the nicest areas and grow them out.
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Feasoghorm

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Lol yeah...we dnt knw.
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TheOffice
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Verum subsequentis, what a coincidence, I just visited your 'isolate thread' 
So coming from someone who has done so much work, Iam satisfied with the answer. I'll just go out and transfer the good parts.
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Inthepit
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Re: The basics of working with Agar [Re: TheOffice] 1
#27307832 - 05/14/21 04:48 AM (3 years, 8 months ago) |
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TheOffice
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Re: The basics of working with Agar [Re: Inthepit]
#27307875 - 05/14/21 05:37 AM (3 years, 8 months ago) |
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Lol, thanks Inthepit.. I did just that, but I had to copy/paste incl. [url... Etc
And to check if it works..
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TheOffice
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Re: The basics of working with Agar [Re: TheOffice]
#27307876 - 05/14/21 05:37 AM (3 years, 8 months ago) |
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Hindsight
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Re: The basics of working with Agar [Re: TheOffice]
#27307984 - 05/14/21 07:40 AM (3 years, 8 months ago) |
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Quote:
TheOffice said: Any advice on how to keep a good culture going?
Just keep the culture on an agar plate in the fridge. You can make multiple agar plates off a single plate by doing transfers. When you go to grain, don't use all the agar in your plate - leave some, and then divide that up into several more plates. Once the plates have grown out, store them in the fridge. They keep a LONG time in the fridge, though slants will last even longer.
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verum subsequentis
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Re: The basics of working with Agar [Re: TheOffice]
#27308043 - 05/14/21 08:36 AM (3 years, 8 months ago) |
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Quote:
TheOffice said: Verum subsequentis, what a coincidence, I just visited your 'isolate thread' 
So coming from someone who has done so much work, Iam satisfied with the answer. I'll just go out and transfer the good parts.

Speaking of coincidence, I just emptied out all those slants last night. Time to repurpose them.
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TheOffice
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Re: The basics of working with Agar [Re: Hindsight]
#27308287 - 05/14/21 11:55 AM (3 years, 8 months ago) |
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Storing them on agar plates sounds more than sufficient for me. I also found a tek which dilutes lc with distilled water for storage. Going to give them both a try
Thanks again for the response!
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TheOffice
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Let's just agree that there is no such thing as coincidence 
Furthermore, I am looking forward to your next journey! Which leads me to a short question: When it comes to agar work/sectoring, a lot of people say " make your agar with less nutrients, so it will stretch out and show more vigerous growth"
Why wouldn't you want the mycelium to stretch out? And just make your standard agar recipe as low on nutrients as possible? Is there a downside to this?
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verum subsequentis
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Re: The basics of working with Agar [Re: TheOffice]
#27308294 - 05/14/21 12:02 PM (3 years, 8 months ago) |
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Be sure to wrap petris nice and tight and put them in an insulated cooler to prevent condensation issues. If done right, petris stored in a fridge can last a good while.
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verum subsequentis
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Re: The basics of working with Agar [Re: TheOffice]
#27308300 - 05/14/21 12:07 PM (3 years, 8 months ago) |
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Quote:
TheOffice said: Let's just agree that there is no such thing as coincidence 
Furthermore, I am looking forward to your next journey! Which leads me to a short question: When it comes to agar work/sectoring, a lot of people say " make your agar with less nutrients, so it will stretch out and show more vigerous growth"
Why wouldn't you want the mycelium to stretch out? And just make your standard agar recipe as low on nutrients as possible? Is there a downside to this?
Honestly, I've done very minimal testing with low nute agar. I've never had a need to really try or care. I do 2% Malt and have great results.
PS... I'm still trying to figure out if there are or are not coincidences. Let's say I wake up and realize that I forgot to take the trash out and the trash truck is coming. I rush out the door bare foot and squish a caterpillar while hurrying to the trash. Meanwhile I realize that I need to have a shit. I rush back in the house and sit on the throne. While excreting I find caterpillar moosh on my foot. I grab a piece of toilet paper and realize it has Podesta's face on it. I use it to clean the caterpillar off my foot and the shite off my arse hole. Now, is it a coincidence that Podesta now has shite and caterpillar moosh on his face, or is there some deeper meaning. I don't know.
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TheOffice
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Quote:
verum subsequentis said:
Quote:
TheOffice said: Let's just agree that there is no such thing as coincidence 
Furthermore, I am looking forward to your next journey! Which leads me to a short question: When it comes to agar work/sectoring, a lot of people say " make your agar with less nutrients, so it will stretch out and show more vigerous growth"
Why wouldn't you want the mycelium to stretch out? And just make your standard agar recipe as low on nutrients as possible? Is there a downside to this?
Honestly, I've done very minimal testing with low nute agar. I've never had a need to really try or care. I do 2% Malt and have great results.
:
PS... I'm still trying to figure out if there are or are not coincidences. Let's say I wake up and realize that I forgot to take the trash out and the trash truck is coming. I rush out the door bare foot and squish a caterpillar while hurrying to the trash. Meanwhile I realize that I need to have a shit. I rush back in the house and sit on the throne. While excreting I find caterpillar moosh on my foot. I grab a piece of toilet paper and realize it has Podesta's face on it. I use it to clean the caterpillar off my foot and the shite off my arse hole. Now, is it a coincidence that Podesta now has shite and caterpillar moosh on his face, or is there some deeper meaning. I don't know.
Remarkable, so many plates done and not really experimented onn the recepy. Never change a winning team 
LOL! Struggling with like minded thoughts.. Melissa McCarthy stands on times Square, a pigeon flies over and drops a shit exactly on that little spot where Melissa isn't standing.. Is that a coincidence or not?
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