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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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Cloning on pastyplates tips?
    #25635285 -

Hey, just got some good looking mushies from my trays and tried cloning a few to agar on pastyplates: https://www.shroomery.org/forums/showflat.php/Number/19208976

Problem is since I had to go at the agar from such a vertical angle I couldn't seen to get the tissue off my scalpel by dragging it. So I may have been a bit unsanitary and scraped it off the blade using the underside of the lid.

Does anyone use pastyplates and has similar issues? Is there an easier way or do I just need to refine my technique?

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Re: Cloning on pastyplates tips? [Re: MrFahrenheit]
    #25635312 -

You could try grabbing the tissue or wedge more 'loosely' so that it's almost falling off when you lift it up.
That way it's more likely it will slip off the scalpel easier! Gl


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Cakes inside Water Tub with optimal Surface Conditions

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Re: Cloning on pastyplates tips? [Re: Mateja]
    #25635328 -

Mateah said:
You could try grabbing the tissue or wedge more 'loosely' so that it's almost falling off when you lift it up.
That way it's more likely it will slip off the scalpel easier! Gl


Dang, I had a hard enough time getting it to stay on the scalpel. Maybe I need a different shaped blade..

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Re: Cloning on pastyplates tips? [Re: MrFahrenheit]
    #25635349 -

practice makes the master my friend. using a new scalpel blade can help as well worn scalpel blades tend to be microscopically serrated and like to hold on to the tissue. also, if you scrape a tissue sample free, then scrape it of on inner stipe tissue, then gingerly scoop it up with the scalpel, then it should come off easier within the plate. also, pouring your plates a little thicker can help because you can submerge the tip of the scalpel blade in the thick agar and force the tissue off.


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Masters and slaves hunting clones        How not to isolate (it worked, but)


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Re: Cloning on pastyplates tips? [Re: verum subsequentis]
    #25635353 -

verum subsequentis said:
practice makes the master my friend. using a new scalpel blade can help as well worn scalpel blades tend to be microscopically serrated and like to hold on to the tissue. also, if you scrape a tissue sample free, then scrape it of on inner stipe tissue, then gingerly scoop it up with the scalpel, then it should come off easier within the plate. also, pouring your plates a little thicker can help because you can submerge the tip of the scalpel blade in the thick agar and force the tissue off.


Both very good ideas, thank you. I'll be getting a new blade and trying this tomorrow. Might try cloning 2 pins to see how that works as well, any experience trying that? Just saw it in the LGM videos.

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Re: Cloning on pastyplates tips? [Re: verum subsequentis]
    #25635414 -

verum subsequentis said:
practice makes the master my friend. using a new scalpel blade can help as well worn scalpel blades tend to be microscopically serrated and like to hold on to the tissue. also, if you scrape a tissue sample free, then scrape it of on inner stipe tissue, then gingerly scoop it up with the scalpel, then it should come off easier within the plate. also, pouring your plates a little thicker can help because you can submerge the tip of the scalpel blade in the thick agar and force the tissue off.


I learn something new here everyday.
:themoreyouknow:


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Re: Cloning on pastyplates tips? [Re: CzM8]
    #25635417 -

i have lots of experience cloning pins. its a great idea. what do you want to know


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Masters and slaves hunting clones        How not to isolate (it worked, but)


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Re: Cloning on pastyplates tips? [Re: MrFahrenheit]
    #25635490 -

If they are tiny pins, you can literally spear it, of flame some tweezers, and put the whole pin into the dish and it will revert back to mycelium within a few days..larger mushrooms you should rip longways and take a small chunk from inside the stem, which isn't usually particularly sticky. Just give the scalpel a tap on the edged of the dish


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"How it should & shouldn't look "      "Recognizing contamination"      "Rye Berry Prep"      "Franks G2G"      "Humidity Chamber"      "Agar Recipes"

"How to dial in your monotubs like a Boss"      "Pastywhyte's Easy Agar Tek"      "Elementary [Damion5050]Coir Tek"        "SGFC"      "SAB Tek"      "Easy AF Shoeboxes"


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Re: Cloning on pastyplates tips? [Re: verum subsequentis]
    #25636140 -

verum subsequentis said:
i have lots of experience cloning pins. its a great idea. what do you want to know


So I just put 3 pins on 3 plates. Ripped the bits of sub off 2 and left the third because I wanted to know if that causes issues. Wasn't very sterile as all I did was iso my gloves and pick it by hand. Figured that it doesn't make sense to be overly cautious since air-exposed tissue will be on agar anyways. Was I wrong to assume that? Also made 4 prints overnight, hoping those are clean as well. Is using scissors to cut tinfoil down to size really frowned upon? The tinfoil piece I used is way oversized at the moment...

Edited by MrFahrenheit (11/24/18 12:00 PM)

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Re: Cloning on pastyplates tips? [Re: MrFahrenheit]
    #25636237 -

not being very sterile when doing sterile work NEVER increases your chances but i get your point there. i'd still encourage you to be as clean as is practical.

alright, here's the deal. what you are attempting will result in an almost guaranteed contam bonanza. if it doesn't then sweet lord is with you... amen... but it will. then it's just you, your sab and your scalpel against the world. how good are you?

what's good is that the myc from the pin is in the business of growing very quickly. this gives you a chance to snatch up clean myc before the contams can catch up. I'd much prefer doing this type of work in actual petri dishes but it can be done in rounds.

check on them often. multiple times a day. be prepared to drop what you are doing and jump into a SAB session when the time strikes. you're going to want to grab some fresh myc once you see it climbing across the agar. if everything is clean (IF IS A BIG WORD) then you can just chill and take a transfer later but I THINK it won't be clean. you gotta snatch that shit up quick and get it to a new plate or two.

the substrate will not help at all and should be removed.

due to the probability of ending up in a battle royal with contams, i typically prefer to take my pins from plates. i thought that that was what you were planning on doing. i like to let pins get bigger when growing in the evil open air so i can take an inner tissue sample and save myself all the hassel.


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Masters and slaves hunting clones        How not to isolate (it worked, but)


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Re: Cloning on pastyplates tips? [Re: verum subsequentis]
    #25636394 -

verum subsequentis said:
not being very sterile when doing sterile work NEVER increases your chances but i get your point there. i'd still encourage you to be as clean as is practical.

alright, here's the deal. what you are attempting will result in an almost guaranteed contam bonanza. if it doesn't then sweet lord is with you... amen... but it will. then it's just you, your sab and your scalpel against the world. how good are you?

what's good is that the myc from the pin is in the business of growing very quickly. this gives you a chance to snatch up clean myc before the contams can catch up. I'd much prefer doing this type of work in actual petri dishes but it can be done in rounds.

check on them often. multiple times a day. be prepared to drop what you are doing and jump into a SAB session when the time strikes. you're going to want to grab some fresh myc once you see it climbing across the agar. if everything is clean (IF IS A BIG WORD) then you can just chill and take a transfer later but I THINK it won't be clean. you gotta snatch that shit up quick and get it to a new plate or two.

the substrate will not help at all and should be removed.

due to the probability of ending up in a battle royal with contams, i typically prefer to take my pins from plates. i thought that that was what you were planning on doing. i like to let pins get bigger when growing in the evil open air so i can take an inner tissue sample and save myself all the hassel.


Pins on plates?! The only time I've had that happen is when I left mine for months without using them. So far I did three inner tissue to agar and three pins to agar. My plan was that at least one of them has to be slightly clean enough to grab a healthy piece or two for transfers. Wish me luck!

Ps: any comment on using scissors to cut spore print tinfoil down to size? I could heat the scissors with my torch before cutting. Shouldn't cause any issues?

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Re: Cloning on pastyplates tips? [Re: MrFahrenheit]
    #25636442 -

it depends on the cleanliness of the scissors. obviously if you had some all metal scissors and pced them, it'd be totally fine. if you iso the scissors up and flame the blades you should be fine.

i don't usually take care to time how long my plates have been sitting around before they pin. i just set them aside after i'm done with them and check the pile every now and then. if i find an nice pin or and incredible "pinset" on a culture, i grab it up and put it on a new plate. unless i already have those genetics rolling strong.


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Masters and slaves hunting clones        How not to isolate (it worked, but)


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Re: Cloning on pastyplates tips? [Re: verum subsequentis]
    #25636469 -

I've had about 5 petris pin in 4-5 weeks after transfer. Some people say that it helps if you take the film off and give them extra fresh air, it didn't seem to matter to my plates.


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Re: Cloning on pastyplates tips? [Re: CzM8]
    #25636478 -

i'd not recommend that. it may help but the beauty of an agar pin is that it is sterile. i guess you could take it off and try to never breath on or around it and keep all currents to a minimum. or unwrap them and leave them in a clean tote. but, why? i just forget about them and go on with life for a while. then i check the stash when i remember. it only takes a second to look through a couple plates.



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Masters and slaves hunting clones        How not to isolate (it worked, but)


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Re: Cloning on pastyplates tips? [Re: verum subsequentis]
    #25636505 -

verum subsequentis said:
i'd not recommend that. it may help but the beauty of an agar pin is that it is sterile. i guess you could take it off and try to never breath on or around it and keep all currents to a minimum. or unwrap them and leave them in a clean tote. but, why? i just forget about them and go on with life for a while. then i check the stash when i remember. it only takes a second to look through a couple plates.




:tweak:
I thought I had a lot of plates. FUAHHHHHHHH


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Re: Cloning on pastyplates tips? [Re: CzM8]
    #25636521 -

i am the master of working way harder than i need too. it's a blessing and a curse.


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Masters and slaves hunting clones        How not to isolate (it worked, but)


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Re: Cloning on pastyplates tips? [Re: verum subsequentis]
    #25636592 -


No really


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Everything you need in one spot.

My Q&A thread. Ask me questions here rather than in Private Message
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Re: Cloning on pastyplates tips? [Re: bodhisatta]
    #25636637 -

You don't ever pitch them? Or is that plates from like a 2 month period?


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Re: Cloning on pastyplates tips? [Re: CzM8]
    #25636768 -

i was literally just being a nerd and letting them build up for a fun picture. no way i'd do that in two months. even in the middle of my super over elaborate isolation i was only using a couple hundred a month


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Masters and slaves hunting clones        How not to isolate (it worked, but)


Edited by verum subsequentis (11/24/18 04:56 PM)

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Re: Cloning on pastyplates tips? [Re: verum subsequentis]
    #25636813 -

Lol GOOD! I just pitch mine after a couple weeks-months. I've really just gotten super into myco in general.

Out of curiosity, when you do transfers looking for better growth. how many transfers do you do off of your nicer plates; not mono's, but say from spores?

I ask this because I keep getting OKAY growth even after T-8+. Im not in search of an isolate, just vigorous and even growth. I've been doing 3-4 transfers per petri dish( so maybe 3-15 per T-value), depending on how my plates are looking.


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