Paradox
©
Fisana

Jump to content


Photo
* * * * * 1 votes

Sectoring / Isolating


  • Please log in to reply
53 replies to this topic

#21 MsBehavin420

MsBehavin420

    Mycotopiate

  • Free Member
  • 921 posts

Posted 02 June 2019 - 08:26 AM

That seems to be the consensus. Man I was hoping for some more science fun

#22 onediadem

onediadem

    Insidious Drivel

  • OG VIP
  • 15,148 posts

Awards Bar:

Posted 02 June 2019 - 03:34 PM

I pour my agar pretty thick, at least half the plate so I can shave off the sections I want. I do that so when I do go to grain, it seems the thicker the agar, the less likely it is to stick to the jar.

 

To answer about storage, I always make up at least twenty plates of the final transfer so I have plenty to work with for a very long time. Then I transfer into sterile distilled vials of water. When you get to that point, make sure every plate is pristine because your going to agitate the top of the myc with a sterile needle filled with the water, squirt some water onto the plate and keep doing that until you have enough in the syringe to transfer into the vial. Repeat for however many you want to store. I keep them in the box they came in in a baggie, stored in the fridge. I have no idea how everyone else does it, and they can chime in also but that is how I do it. When I run out I start over.


  • bezevo and MsBehavin420 like this

#23 Taproot

Taproot

    Mycophiliac

  • Free Member
  • 34 posts

Posted 16 June 2019 - 11:41 PM

Sorry, and yes. Transfer those sections that (Small sections like no bigger than 1/4 inch square) that I explained above. I prefer to just get that one strand. Once your strand grows out on the plate, get it to grain. Then knock up your substrate. When it fruits, I myself do not mess with anything less that a 5 mushroom cluster to clone on the first generation. To me, anything less is not worth the time or agar and plates. There is an end goal here. Repeat that process 3-4 times. I have done both, and rarely have I ever had a problem. The last generation Is the golden egg. I put that to agar, let it grow out and make vials of sterile water water to put that last generation into. 
 
As I said, it is a very long process. But oh so worth it. Figure minimum of two months per generation, if you have no contam issues. I always wait until the second or third flush after a dunk also. I don't know why second flushes are always the best for me, but they are.
 
I dont know if anyone else does this with their agar, but I always pour two sleeves at a time. I use 1 inch rolls cut from saran wrap to seal the plates so they don't dry out. If they contam, they are not even opened. They last me a few months, and I am not repouring, or leaving a sleeve open. I refuse to use plates from an open sleeve once they have been removed from the glovebox.
 
Anyway, hope that helps and sorry for being so chatty today.


Im starting to get further a long in this process and have two questions. First, do you take clones from a 5 mushroom cluster from the very first flush of that tub? Does it matter what flush you take it from? And by "generation"are you reffering to the cycle of clone to agar, agar to grain, grain to substrate then that first flush as generation #1 then repeating that and then the next cycle of that is generation two?

#24 crazy1

crazy1

    Celestial Traveler

  • VIP
  • 1,998 posts

Posted 17 June 2019 - 06:43 AM

One is saying she'll take a 5 shroom cluster off of the first flush if cloning.

Then she mentions the final flush as the golden egg.

 

IF you go agar to agar, grain to grain, you should keep track of what transfer/generation you are on. The mycelium will senesce after ???? many transfers.

 

Peace


Edited by crazy1, 17 June 2019 - 06:44 AM.


#25 Taproot

Taproot

    Mycophiliac

  • Free Member
  • 34 posts

Posted 17 June 2019 - 07:58 AM

One is saying she'll take a 5 shroom cluster off of the first flush if cloning.
Then she mentions the final flush as the golden egg.
 
IF you go agar to agar, grain to grain, you should keep track of what transfer/generation you are on. The mycelium will senesce after ???? many transfers.
 
Peace

I'm sorry but I need to clarify again, is a generation each flush you get from colonized substrate or is it each time you do a cycle of agar to grain to substrate to fruit? And by golden egg she means it's best to instead wait until the final flush and clone that cluster instead of the preceding ones? Or does she mean clone a 5 mushroom cluster first flush and then after you've done that and used that clone for a monotub or whatever, the final flush from it will be the golden egg (a really good flush)?Thanks!

#26 Taproot

Taproot

    Mycophiliac

  • Free Member
  • 34 posts

Posted 17 June 2019 - 08:18 AM

In case anyone is curious how the transfers are going here are some pictures. The individual plates are the second agar transfers (A.T.#2) and the group picture is of the third transfers (A.T.#3) I just transferred those again yesterday (A.T.#4). I will probably use the A.T.#4's to spawn a monotub for testing.

Attached Thumbnails

  • 20190531_202520.jpg
  • 20190531_202458.jpg
  • 20190531_202447.jpg
  • 20190531_202428.jpg
  • 20190531_202413.jpg
  • 20190616_161522.jpg


#27 Misfit

Misfit

    Mycotopiate

  • VIP
  • 428 posts

Posted 17 June 2019 - 08:35 AM

Still learning about isolation. What is it about plate E #2 that looked promising. All the others I totally get. Just not sure what it was about that one.

#28 CatsAndBats

CatsAndBats

    this motherfucker

  • OG VIP
  • 11,605 posts

Awards Bar:

Posted 17 June 2019 - 12:19 PM

 

Is there any recommended posts I can read that will help me understand what the "strongest looking sector" would look like? I'm pretty new to this and am still unsure. I was hoping to get a experienced member to show me what they would do / where they would sector it.

https://mycotopia.ne...k-some-agar-30/

 

 

Thanks Mikey.

 

Op, this is my simpler agar thread:

 

https://mycotopia.ne...izes-agar-agar/

 

 

Definitely cannot go wrong using Cat's thread. Awesome good info in there and hey, it's got lots of pics too. Good call Mikey. I admit I was much too lazy to dig for that one lol..

You don't have it as your start up page in your browser?!

 

Hell's Bell's and Cockle Shells!

 

I really appreciate this thread and the input given!

 

I'm excited to work on some agar skills after seeing the suggestions from one and coors!

Hi Roc!

 

I have two follow up questions for the pro's. First, how do you go about producing your 3rd or 4th cluster that you then clone? Do you clone/agar/sector/fruit/clone/agar/sector/fruit until your at your fourth cluster? I was told by some that senescence would be an issue if I didn't use spores from said clones after fruiting/cloning/sectoring/fruiting multiple times. So I guess my question is how many times can I do this cycle before I experience senescence and then have to use spores from my clones. Or do I? The second question, is when you sector a piece of desirable mycelium, do you take the individual ropey rhizomorphic piece and put it on another plate or do you cut a, for example 3mm x 3mm chuck of agar with the desired mycelium and others and place that on a fresh plate. I was worried with some of the sectors I took that I took too little mycelium. Sorry if this is too much to ask I'm still figuring out what i can and should be asking and what is overkill or frowned upon. Once again thank you all for the tips and support / energy invested in your posts. It means the world to me. I'll upload pictures of my work tomorrow for you to see what I've done.

I just take wisps of myc from where it most aggressively grows with a scalpel and then slice it into new agar.

 

Can you please simplify your other question or throw some paragraph breaks in there? Old men with ADHD can't read too alright. :biggrin:

 

 

One is saying she'll take a 5 shroom cluster off of the first flush if cloning.
Then she mentions the final flush as the golden egg.
 
IF you go agar to agar, grain to grain, you should keep track of what transfer/generation you are on. The mycelium will senesce after ???? many transfers.
 
Peace

I'm sorry but I need to clarify again, is a generation each flush you get from colonized substrate or is it each time you do a cycle of agar to grain to substrate to fruit? And by golden egg she means it's best to instead wait until the final flush and clone that cluster instead of the preceding ones? Or does she mean clone a 5 mushroom cluster first flush and then after you've done that and used that clone for a monotub or whatever, the final flush from it will be the golden egg (a really good flush)?Thanks!

 

 

 

Generation is spore to spore, in other words an entire life cycle. Transfer is from plate to plate.


  • roc, crazy1 and Taproot like this

#29 roc

roc

    Forum International Moderator

  • Moderator
  • 7,928 posts

Awards Bar:

Posted 17 June 2019 - 12:35 PM

https://mycotopia.ne...izes-agar-agar/

 

 

 

Hell's Bell's and Cockle Shells!

 

I really appreciate this thread and the input given!

 

I'm excited to work on some agar skills after seeing the suggestions from one and coors!

Hi Roc!

 

Hello!

Nice link!


  • CatsAndBats likes this

#30 HrVanker

HrVanker

    Mycophiliac

  • Free Member
  • 323 posts

Awards Bar:

Posted 17 June 2019 - 06:41 PM

I have yet to graduate to agar. But it will happen eventually.

One idea I read long ago, was Anne Halonium's use of square PP5 bottles for her final transfers. She installed injection ports, and poured the agar on the side of the bottle. Most tools used for transfer can fit through the neck.

The primary purpose for this was to inject with sterile water to make a slurry. No open jars to transfer agar wedges, so it should cut down on contam rates. You could do a lot of jars from one bottle.

I know that Anne is banned here. But I think she had a couple good things going for her.

Sent from my SM-G960U using Tapatalk

#31 Taproot

Taproot

    Mycophiliac

  • Free Member
  • 34 posts

Posted 17 June 2019 - 06:44 PM

Sorry, and yes. Transfer those sections that (Small sections like no bigger than 1/4 inch square) that I explained above. I prefer to just get that one strand. Once your strand grows out on the plate, get it to grain. Then knock up your substrate. When it fruits, I myself do not mess with anything less that a 5 mushroom cluster to clone on the first generation. To me, anything less is not worth the time or agar and plates. There is an end goal here. Repeat that process 3-4 times. I have done both, and rarely have I ever had a problem. The last generation Is the golden egg. I put that to agar, let it grow out and make vials of sterile water water to put that last generation into.

As I said, it is a very long process. But oh so worth it. Figure minimum of two months per generation, if you have no contam issues. I always wait until the second or third flush after a dunk also. I don't know why second flushes are always the best for me, but they are.

I dont know if anyone else does this with their agar, but I always pour two sleeves at a time. I use 1 inch rolls cut from saran wrap to seal the plates so they don't dry out. If they contam, they are not even opened. They last me a few months, and I am not repouring, or leaving a sleeve open. I refuse to use plates from an open sleeve once they have been removed from the glovebox.

Anyway, hope that helps and sorry for being so chatty today.


Im starting to get further a long in this process and have two questions. First, do you take clones from a 5 mushroom cluster from the very first flush of that tub? Does it matter what flush you take it from? And by "generation"are you reffering to the cycle of clone to agar, agar to grain, grain to substrate then that first flush as generation #1 then repeating that and then the next cycle of that is generation two?

Is there any recommended posts I can read that will help me understand what the "strongest looking sector" would look like? I'm pretty new to this and am still unsure. I was hoping to get a experienced member to show me what they would do / where they would sector it.

https://mycotopia.ne...k-some-agar-30/
Thanks Mikey.

Op, this is my simpler agar thread:

https://mycotopia.ne...izes-agar-agar/

Definitely cannot go wrong using Cat's thread. Awesome good info in there and hey, it's got lots of pics too. Good call Mikey. I admit I was much too lazy to dig for that one lol..

You don't have it as your start up page in your browser?!

Hell's Bell's and Cockle Shells!

I really appreciate this thread and the input given!

I'm excited to work on some agar skills after seeing the suggestions from one and coors!

Hi Roc!

I have two follow up questions for the pro's. First, how do you go about producing your 3rd or 4th cluster that you then clone? Do you clone/agar/sector/fruit/clone/agar/sector/fruit until your at your fourth cluster? I was told by some that senescence would be an issue if I didn't use spores from said clones after fruiting/cloning/sectoring/fruiting multiple times. So I guess my question is how many times can I do this cycle before I experience senescence and then have to use spores from my clones. Or do I? The second question, is when you sector a piece of desirable mycelium, do you take the individual ropey rhizomorphic piece and put it on another plate or do you cut a, for example 3mm x 3mm chuck of agar with the desired mycelium and others and place that on a fresh plate. I was worried with some of the sectors I took that I took too little mycelium. Sorry if this is too much to ask I'm still figuring out what i can and should be asking and what is overkill or frowned upon. Once again thank you all for the tips and support / energy invested in your posts. It means the world to me. I'll upload pictures of my work tomorrow for you to see what I've done.

I just take wisps of myc from where it most aggressively grows with a scalpel and then slice it into new agar.

Can you please simplify your other question or throw some paragraph breaks in there? Old men with ADHD can't read too alright. :biggrin:

One is saying she'll take a 5 shroom cluster off of the first flush if cloning.
Then she mentions the final flush as the golden egg.

IF you go agar to agar, grain to grain, you should keep track of what transfer/generation you are on. The mycelium will senesce after ???? many transfers.

Peace

I'm sorry but I need to clarify again, is a generation each flush you get from colonized substrate or is it each time you do a cycle of agar to grain to substrate to fruit? And by golden egg she means it's best to instead wait until the final flush and clone that cluster instead of the preceding ones? Or does she mean clone a 5 mushroom cluster first flush and then after you've done that and used that clone for a monotub or whatever, the final flush from it will be the golden egg (a really good flush)?Thanks!

Generation is spore to spore, in other words an entire life cycle. Transfer is from plate to plate.
Ok I finally think I understand. I have one more question. "Insidious drivel" said to clone a 5 mushroom cluster and repeat this process 3-4 times and that the last generation is the golden egg. Yet "This motherfucker" said that "generation meant from spore to spore"

So does this mean I'm cloning mushrooms each cluster or am I collecting spores from each cluster from succesive generations, starting them and then collecting spores from the next 5 mushroom cluster ect.? Im so confused... Does anyone see what I'm saying?

I just cloned a pin from this 6 cluster in a tub on its first flush. Now im confused about if i was just supposed to get a print from them.

Attached Thumbnails

  • 20190617_184901.jpg

Edited by Taproot, 17 June 2019 - 06:47 PM.


#32 HrVanker

HrVanker

    Mycophiliac

  • Free Member
  • 323 posts

Awards Bar:

Posted 17 June 2019 - 06:53 PM

Ok I finally think I understand. I have one more question. "Insidious drivel" said to clone a 5 mushroom cluster and repeat this process 3-4 times and that the last generation is the golden egg. Yet "This motherfucker" said that "generation meant from spore to spore"

So does this mean I'm cloning mushrooms each cluster or am I collecting spores from each cluster from succesive generations, starting them and then collecting spores from the next 5 mushroom cluster ect.? Im so confused... Does anyone see what I'm saying?

Take a sample from a 5 shroom cluster, throw it on agar.

Once that has grown out, and you find nice healthy strand of myc outgrowing everything else, take a *small* sample from it, as someone else said, 1/4" square (about the size of an LSD blotter). Transfer that specimen to a new plate and grow it out.

Once the second plate is covered in healthy growth, take samples and inoculate grain jars with it.

Use those jars for a tub or however you want to fruit it.

When fruiting the sub that was prepared with the grain that you 'nocced up with the agar, find a healthy 5 mushroom cluster. Take a sample from the cluster, and put it on agar.

You have now completed one generation and started the next.

Edit: If you weren't taking clones, and just doing spore prints. One generation would be from the spore print/syringe you start with, through the printing stage of the shrooms from said spores. When you start a grow with the spores from that first grow, you are on generation 2.

Edited by HrVanker, 17 June 2019 - 06:58 PM.

  • Taproot likes this

#33 Taproot

Taproot

    Mycophiliac

  • Free Member
  • 34 posts

Posted 17 June 2019 - 07:00 PM

Oh okay. I thought the previous posters meant to say i needed to collect spores each time. So much conflicting information out there and it confuses the heck out of me. I know everyone has a different way of doing things though. Not one way better than the other just a different way of arriving at the same destination. I'm glad to know I can clone 5 generations without worrying about senescence!
  • HrVanker likes this

#34 jkdeth

jkdeth

    Mycotopiate

  • Free Member
  • 2,472 posts

Awards Bar:

Posted 17 June 2019 - 07:16 PM

Do it whichever way you want. Otherwise we just end up arguing semantics and which name to call things.

There is a limit. I can't tell you exactly what it is. Young cultures out perform old cultures. Succesive cloning ages the culture. Starting from spore resets it.
  • Microbe and Taproot like this

#35 onediadem

onediadem

    Insidious Drivel

  • OG VIP
  • 15,148 posts

Awards Bar:

Posted 17 June 2019 - 10:09 PM

 

Sorry, and yes. Transfer those sections that (Small sections like no bigger than 1/4 inch square) that I explained above. I prefer to just get that one strand. Once your strand grows out on the plate, get it to grain. Then knock up your substrate. When it fruits, I myself do not mess with anything less that a 5 mushroom cluster to clone on the first generation. To me, anything less is not worth the time or agar and plates. There is an end goal here. Repeat that process 3-4 times. I have done both, and rarely have I ever had a problem. The last generation Is the golden egg. I put that to agar, let it grow out and make vials of sterile water water to put that last generation into. 
 
As I said, it is a very long process. But oh so worth it. Figure minimum of two months per generation, if you have no contam issues. I always wait until the second or third flush after a dunk also. I don't know why second flushes are always the best for me, but they are.
 
I dont know if anyone else does this with their agar, but I always pour two sleeves at a time. I use 1 inch rolls cut from saran wrap to seal the plates so they don't dry out. If they contam, they are not even opened. They last me a few months, and I am not repouring, or leaving a sleeve open. I refuse to use plates from an open sleeve once they have been removed from the glovebox.
 
Anyway, hope that helps and sorry for being so chatty today.


Im starting to get further a long in this process and have two questions. First, do you take clones from a 5 mushroom cluster from the very first flush of that tub? Does it matter what flush you take it from? And by "generation"are you reffering to the cycle of clone to agar, agar to grain, grain to substrate then that first flush as generation #1 then repeating that and then the next cycle of that is generation two?

 

 

A generation is the complete cycle, so yes. It doesnt matter which flush you take the clone from, but I myself prefer the second flush because the mushrooms seem to be the best IMO. After your first clone, you want the cluster to be the biggest of the group. Each generation will throw larger and larger clusters. That is why I never clone a cluster with less than 5-6 mushrooms on the first clone. The second gen you should have clusters with at least 8-10 mushrooms. My third gen clusters never have less than 20-30 mushrooms per cluster. If I see no degrading, I go for a fourth. The nice thing about getting to a third or fourth gen clone is there is no guessing what your substrate will do. They are also so prolific that they wipe out the nutes within 2 to sometimes 3 flushes and you are not spending time and space on trying to get a few mushrooms on later flushes. The more flushes you try to milk out of a substrate, the chances of contamination grow exponentially.


  • crazy1, Microbe, peacefrog and 2 others like this

#36 Taproot

Taproot

    Mycophiliac

  • Free Member
  • 34 posts

Posted 18 June 2019 - 12:43 AM

After reading through this a second time it seems like the wisest thing to do would to collect spores from each cluster instead of clones to prevent senescence.

As Mycotopiate said, "there is a limit"

Now the big question is, does collecting spores from large clusters each generation yield the same results as cloning clusters from each generation? If so I'm not sure why anyone would do anything but collect the spores since it would be the best way to ensure the healthiest most vigorous young mycelium time after time instead of using a 5th generation clone that's been alive for a year.

#37 HrVanker

HrVanker

    Mycophiliac

  • Free Member
  • 323 posts

Awards Bar:

Posted 18 June 2019 - 12:49 AM

After reading through this a second time it seems like the wisest thing to do would to collect spores from each cluster instead of clones to prevent senescence.

As Mycotopiate said, "there is a limit"

Now the big question is, does collecting spores from large clusters each generation yield the same results as cloning clusters from each generation? If so I'm not sure why anyone would do anything but collect the spores since it would be the best way to ensure the healthiest most vigorous young mycelium time after time instead of using a 5th generation clone that's been alive for a year.

Do both. But rely on clones for the bulk of your growing. Reserve spores for use when senescence is too far advanced, or you want to start something new for fun.

Using prints and spore syringes does not guarantee any traits will be passed. One mushroom can put out SOOOO many spores that the genetic possibilities are too varied.

Sent from my SM-G960U using Tapatalk
  • Taproot likes this

#38 jkdeth

jkdeth

    Mycotopiate

  • Free Member
  • 2,472 posts

Awards Bar:

Posted 18 June 2019 - 02:22 AM

Cloning, then printing, slowly improves the probability of a better grow from spores. It can take several generations to make a trait or phenotype dominant.

No I'm afraid a grow from a clone print won't be the same as the clone grow. But it will be a little bit closer. Do that 4 to 6 times, you'll make a noticeable improvement in the multispore growth.
  • HrVanker, Microbe, peacefrog and 1 other like this

#39 jkdeth

jkdeth

    Mycotopiate

  • Free Member
  • 2,472 posts

Awards Bar:

Posted 18 June 2019 - 02:45 AM

So probably just getting more confusing. It will more sense with the hands on. Actually seeing the grows.

#40 Taproot

Taproot

    Mycophiliac

  • Free Member
  • 34 posts

Posted 18 June 2019 - 09:50 AM

You're all amazing. Thank you for the advice. Everyone who has posted has shared their experience and opinions and it's given me a lot of insight. What a wonderful community this is :)
  • roc, crazy1, Tenderfoot and 1 other like this




Like Mycotopia? Become a member today!