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OfflineShr00merN00ber
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First time will attempt cloning
    #26387985 - 12/18/19 01:16 AM (4 years, 1 month ago)

Hey guys, I recently got some growkits that are growing. I just got them because its much faster than starting out with spores and the genetics from this company are usually excellent. This time around I want to take a few tissue samples and grow them out on agar. Here is their current state:

Kit 1







Kit 2



I have three questions:

1) I'm in a dilemma. I'm not sure which one to pick for cloning. In 1st pic of kit 1, the largest and first/fastest mushroom to grow is on the bottom left side. Its the biggest one of them all atm. Both in girth and in height. On the other hand, there is a nice healthy looking cluster on the right side. Would you choose a tissue sample from a cluster or go with the biggest mushroom which pinned first, even though its surrounding genetics are not as good as the cluster?

2) In pic 3 of kit 1, on the left side, you can see many small knots. Will these all grow out to form shrooms or will they not all make it?

3) The grow kit instructions say to water the sides of the bag every day when the first mushrooms appear. I haven't put one drop of water from the day I received it, its been 10 days now. I think it's fine as there is an excess of water to the point of drops sliding down the sides and pins have formed and are growing. Does the sub look too dry or something to any of you?

Thanks for any input!


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Re: First time will attempt cloning [Re: Shr00merN00ber]
    #26388038 - 12/18/19 03:06 AM (4 years, 1 month ago)

1.  That’s easy.. clone both/all the ones you want

2.  Hard to say.. no one is going to be able to give you a definitive answer... if the conditions are right, a good amount of them will.  But odds are, not all will... the myc is kind of hedging it’s bets

3.  Nope.. doesn’t appear to be too dry at all.. doesn’t even appear to be be pulling away from the sides due to shrinkage which is pretty common.  No bluing either, I’d say you’re golden.

Faht


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Re: First time will attempt cloning [Re: Shr00merN00ber]
    #26388045 - 12/18/19 03:18 AM (4 years, 1 month ago)

Quote:

Shr00merN00ber said:
Hey guys, I recently got some growkits that are growing. I just got them because its much faster than starting out with spores and the genetics from this company are usually excellent. This time around I want to take a few tissue samples and grow them out on agar. Here is their current state:

Kit 1







Kit 2



I have three questions:

1) I'm in a dilemma. I'm not sure which one to pick for cloning. In 1st pic of kit 1, the largest and first/fastest mushroom to grow is on the bottom left side. Its the biggest one of them all atm. Both in girth and in height. On the other hand, there is a nice healthy looking cluster on the right side. Would you choose a tissue sample from a cluster or go with the biggest mushroom which pinned first, even though its surrounding genetics are not as good as the cluster?

2) In pic 3 of kit 1, on the left side, you can see many small knots. Will these all grow out to form shrooms or will they not all make it?

3) The grow kit instructions say to water the sides of the bag every day when the first mushrooms appear. I haven't put one drop of water from the day I received it, its been 10 days now. I think it's fine as there is an excess of water to the point of drops sliding down the sides and pins have formed and are growing. Does the sub look too dry or something to any of you?

Thanks for any input!





Hey man! I did the same thing, going from a kit to bulk...


Take samples from some of the shrooms in the cluster, you want that clustering clone :smile:)


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OfflineShr00merN00ber
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Re: First time will attempt cloning [Re: mushhiehunter]
    #26388083 - 12/18/19 04:15 AM (4 years, 1 month ago)

Thanks for the reply guys, can't wait to get started on the real grow!


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OfflineShr00merN00ber
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Re: First time will attempt cloning [Re: Shr00merN00ber]
    #26389856 - 12/19/19 12:13 AM (4 years, 1 month ago)

Update:

Kit 1







Kit 2




Anyone know why the biggest mushroom's cap split like this? Now it makes me feel like I shouldn't clone that one. Things with kit 2 are looking kinda bad. Can any one provide some input? Thanks!


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OfflineShr00merN00ber
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Re: First time will attempt cloning [Re: Shr00merN00ber]
    #26390245 - 12/19/19 08:03 AM (4 years, 1 month ago)

Update:

Just got back from work and the 'X' shaped cap splitting is getting even more pronounced and I would really like some advice on what to do. Should I pick it now or leave it? The gills look like they have not developed spores, yet it is bruising blue at the same time. I can see a type of 'veil' going down the stem of the mushroom but in my nooby opinion, this is not the veil cap that broke. Could someone more experienced take a look and let me know what to do?

Also, related to my main aim which is to clone, maybe this big fat (mutant?) might not be the way to go. I'm uploading some pic so you can see, the surrounding mushrooms look strange too.



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Re: First time will attempt cloning [Re: Shr00merN00ber]
    #26390258 - 12/19/19 08:20 AM (4 years, 1 month ago)

Usually splitting like that means the fruits either dry (which they don’t look dry) or the mushroom grew so fast that the fruit couldn’t keep up with itself and split... at least when caps split at the edge that’s why.  Your probably ok to clone it, I don’t think it’s infected or anything.

I’d clone any of them.

Maybe someone else has a different/better answer. 

Faht


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OfflineShr00merN00ber
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Re: First time will attempt cloning [Re: fahtster]
    #26390261 - 12/19/19 08:25 AM (4 years, 1 month ago)

Cheers Faht for the answer. What about that veil going down the stem? I was reading up about that 'growth spurt' theory. I was also thinking pseudomonas but no clue. Would you harvest that one now or wait?


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Re: First time will attempt cloning [Re: Shr00merN00ber]
    #26390275 - 12/19/19 08:37 AM (4 years, 1 month ago)

I’d take it now.. how deep is the substrate?

Veils do different things.. no need to worry there.

I don’t see any blotching.. maybe don’t clone the one with the white tipped cap.

But shit, if you have the means, I’d clone them all (one from each cluster if clustered) and grow them out and have fun with it.  Keep it fun and you’ll never have a failure, only fun learning experiences.

Faht


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OfflineShr00merN00ber
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Re: First time will attempt cloning [Re: fahtster]
    #26390288 - 12/19/19 08:42 AM (4 years, 1 month ago)

So true Faht, if it ain't fun, it shouldn't be done!


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Re: First time will attempt cloning [Re: fahtster]
    #26390302 - 12/19/19 08:47 AM (4 years, 1 month ago)

Quote:

fahtster said:
I’d take it now.. how deep is the substrate?

Veils do different things.. no need to worry there.

I don’t see any blotching.. maybe don’t clone the one with the white tipped cap.

But shit, if you have the means, I’d clone them all (one from each cluster if clustered) and grow them out and have fun with it.  Keep it fun and you’ll never have a failure, only fun learning experiences.

Faht




Does taking a clone from a fruit in a cluster really make that big of a difference when you grow it out?


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Re: First time will attempt cloning [Re: SpunkyMonkey88]
    #26390316 - 12/19/19 08:59 AM (4 years, 1 month ago)

Not all sub strains perform the same... depends on what you’re looking I suppose... but that’s I say clone them all and see what happens.

Faht


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OfflineShr00merN00ber
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Re: First time will attempt cloning [Re: fahtster]
    #26390329 - 12/19/19 09:11 AM (4 years, 1 month ago)

I made my first attempt on some Pastyplates yesterday, to be ready for cloning. I made 6 plates in total, 2 have excess water that has formed a little pool, I tilted one so that you can see the extent of it. Do I discard the ones with excess water? 10g agar 10g LME 500 ml water.



Edited by Shr00merN00ber (12/19/19 09:12 AM)


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Re: First time will attempt cloning [Re: Shr00merN00ber]
    #26390343 - 12/19/19 09:21 AM (4 years, 1 month ago)

That ones for someone else with pasty plate experience

Faht


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Re: First time will attempt cloning [Re: fahtster]
    #26390361 - 12/19/19 09:32 AM (4 years, 1 month ago)

sometimes i get a little bit of water on top too, you can just turn it upside down and dump it inside the sab before you use it, or not, i’ve left water in there and it was fine :shrug:


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Re: First time will attempt cloning [Re: Shr00merN00ber]
    #26390394 - 12/19/19 09:57 AM (4 years, 1 month ago)

Quote:

Shr00merN00ber said:
I made my first attempt on some Pastyplates yesterday, to be ready for cloning. I made 6 plates in total, 2 have excess water that has formed a little pool, I tilted one so that you can see the extent of it. Do I discard the ones with excess water? 10g agar 10g LME 500 ml water.







Ya dude just dump out the water on your SAB floor next time you use them, if you did it right the water is sterile anyway.

Also once you have a culture that's grown out a bit it usually "eats" up any left over surface water that you couldn't dump..

I get some pooling even in my petris that I pour especially if I pour them on the hot side... it's not a big deal just make sure they stand upright, dont want the water leaking out around the lid... dont know if that could actally happen but if it did it could be a wick for contamination.

That looks like some thick ass agar btw, I made that mistake my first time too. Itll work but it's a bitch to work with.


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Re: First time will attempt cloning [Re: fahtster]
    #26390398 - 12/19/19 09:59 AM (4 years, 1 month ago)

Quote:

fahtster said:
Not all sub strains perform the same... depends on what you’re looking I suppose... but that’s I say clone them all and see what happens.

Faht




But can you still get a nice flush from cloning a fat "lone ranger"?

Or will it generate sparse fruits?


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OfflineShr00merN00ber
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Re: First time will attempt cloning [Re: SpunkyMonkey88]
    #26390412 - 12/19/19 10:10 AM (4 years, 1 month ago)

This point exactly was just on my mind.


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Re: First time will attempt cloning [Re: Shr00merN00ber]
    #26390457 - 12/19/19 10:41 AM (4 years, 1 month ago)

Sure.. the difference probably isn’t crazy substantial if a variety is prone to clustering etc anyway and they can definitely make up the difference in size if there’s fewer.  But if you have a preference, do that... it’s all up to you.. it can make a big difference or it might not also

Faht


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Re: First time will attempt cloning [Re: Shr00merN00ber]
    #26390474 - 12/19/19 10:51 AM (4 years, 1 month ago)

Quote:

Shr00merN00ber said:
This point exactly was just on my mind.



What point?


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Re: First time will attempt cloning [Re: SpunkyMonkey88]
    #26390502 - 12/19/19 11:08 AM (4 years, 1 month ago)

I think your lone vs cluster fruit question.

I’d def clone that big one. It looks cool so if it happens to be a mutation it might show up again and if not it’s obviously got good genes for growth.

I haven’t done pasty plates but that looks like enough water to warrant a draining.


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Re: First time will attempt cloning [Re: A.k.a]
    #26390711 - 12/19/19 01:14 PM (4 years, 1 month ago)

Ok gotcha just wasn't sure if be was referencing me or someone else...


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OfflineShr00merN00ber
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Re: First time will attempt cloning [Re: SpunkyMonkey88] * 1
    #26390817 - 12/19/19 02:33 PM (4 years, 1 month ago)

The same question you had above "can you still get a nice flush from cloning a fat "lone ranger"".

I'm gonna take a few samples tomorrow morning, and I'll take some separately from the cluster ones and the big one and make an experiment myself. I'll keep a log with progress on this thread. Lets see in practice. (Its my first time attempting this and first go at agar, sab, cloning etc.)


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Re: First time will attempt cloning [Re: Shr00merN00ber]
    #26390839 - 12/19/19 02:55 PM (4 years, 1 month ago)

curious myself


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Re: First time will attempt cloning [Re: Shr00merN00ber]
    #26390850 - 12/19/19 03:02 PM (4 years, 1 month ago)

Quote:

Shr00merN00ber said:
The same question you had above "can you still get a nice flush from cloning a fat "lone ranger"".

I'm gonna take a few samples tomorrow morning, and I'll take some separately from the cluster ones and the big one and make an experiment myself. I'll keep a log with progress on this thread. Lets see in practice. (Its my first time attempting this and first go at agar, sab, cloning etc.)




Ok gotcha...hope your prepared to have a shit ton of plates and be ready to make weekly transfers... basically those plates keep adding up, quickly too.

Dont be afraid to dump plates. I've only been playing around with 2 strains (well three but 1 wasn't ever put to grain) and I only made it to T3s and I've probably dumped 30 plates and am sitting on 15 or so... and that's actually pretty good.

So if you plan on making a bunch of clones I hope you got a ton of plates on hand cus ull need em.

Definitely let us know about the clones though...i never see any threads outlining grows that were from clone cultures, or I'm not paying enough attention...


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Re: First time will attempt cloning [Re: SpunkyMonkey88]
    #26390942 - 12/19/19 04:05 PM (4 years, 1 month ago)

I just put a clone plate from a big solitary fruit to grain two days ago. The myc jumped off real quick so off to a good start. My first grow from agar and first clone also.


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Re: First time will attempt cloning [Re: A.k.a]
    #26391442 - 12/19/19 08:17 PM (4 years, 1 month ago)


Quote:

A.k.a said:
I just put a clone plate from a big solitary fruit to grain two days ago. The myc jumped off real quick so off to a good start. My first grow from agar and first clone also.







Edited by SpunkyMonkey88 (12/19/19 08:18 PM)


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Re: First time will attempt cloning [Re: SpunkyMonkey88]
    #26391797 - 12/20/19 02:10 AM (4 years, 1 month ago)

So I gave it a shot this morning and it was generally harder than what I thought. Working in the sab can be annoying and I was sweating quite a lot towards the end, which made my mind go blank and I'm not 100% sure of which transfers went to which plate in the end!! I'm 80% sure... Dammit
Here are some photos







This guy should be safe to eat right?







Argh I'm so pissed at myself for not keeping 100% track of which specimens were going where.. Also one of my agar plates had the consistency of a slushy when I opened it in the SAB, and another just fell out on the base when I tried to tip some water out. Only 4 transfers in total. Will update in a few days.!


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Re: First time will attempt cloning [Re: Shr00merN00ber]
    #26392182 - 12/20/19 09:46 AM (4 years, 1 month ago)

Lol good luck. I see you had the same patience for melting through the plastic to make arm holes as I did.

I hope that cap is genetic.


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OfflineShr00merN00ber
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Re: First time will attempt cloning [Re: A.k.a]
    #26392185 - 12/20/19 09:50 AM (4 years, 1 month ago)

Haha, I wish its not genetic! Its safe to eat though right? Now thats its dried it looks ok but when I look at that photo.. hmmm :confused:


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Re: First time will attempt cloning [Re: Shr00merN00ber]
    #26392200 - 12/20/19 10:00 AM (4 years, 1 month ago)

I meant the split cap I like that one for some reason. I usually get a couple with that weird ball on top with golden teachers.  all of them should be fine to eat.


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Re: First time will attempt cloning [Re: Shr00merN00ber]
    #26392202 - 12/20/19 10:01 AM (4 years, 1 month ago)

It's a mushroom nipple


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OfflineShr00merN00ber
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Re: First time will attempt cloning [Re: SpunkyMonkey88]
    #26392218 - 12/20/19 10:09 AM (4 years, 1 month ago)

:ohshi:


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OfflineShr00merN00ber
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Re: First time will attempt cloning [Re: Shr00merN00ber]
    #26398870 - 12/24/19 09:20 AM (4 years, 1 month ago)

Hey guys update. Here are the plates 4 days later. Out of the 4 I had, I knocked one on the floor and the tissue sample went on the side and hasn't really recovered. Here are the three that are going:

Plate 1 - (There are actually 2 tissue samples in this plate, pretty close to each other, the sample split when I was trying to move it to the center...!) 





Plate 2



Plate 3




Plates 1 & 2 should be the clones from the big boy, plate 3 from the cluster. Plate 2 looks more white irl, but not as much as plate 1.

So my question, do I wait a few more days for the myc to grow out or shall I transfer asap?

I'm just waiting for my pc to cool and will attempt my first go at pouring petris. Wish me luck!


Edited by Shr00merN00ber (12/24/19 09:22 AM)


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Re: First time will attempt cloning [Re: Shr00merN00ber]
    #26398900 - 12/24/19 09:32 AM (4 years, 1 month ago)

Quote:

Shr00merN00ber said:
Hey guys update. Here are the plates 4 days later. Out of the 4 I had, I knocked one on the floor and the tissue sample went on the side and hasn't really recovered. Here are the three that are going:

Plate 1 - (There are actually 2 tissue samples in this plate, pretty close to each other, the sample split when I was trying to move it to the center...!) 





Plate 2



Plate 3




Plates 1 & 2 should be the clones from the big boy, plate 3 from the cluster. Plate 2 looks more white irl, but not as much as plate 1.

So my question, do I wait a few more days for the myc to grow out or shall I transfer asap?

I'm just waiting for my pc to cool and will attempt my first go at pouring petris. Wish me luck!




Personally I would wait a day or so...

My theory is after its established s bit more the outer edges will be that much further away from any contaminated tissue...
Usually something like this for me:


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Re: First time will attempt cloning [Re: SpunkyMonkey88]
    #26398905 - 12/24/19 09:38 AM (4 years, 1 month ago)

Thanks so much Spunky for your help in all of this. I was wondering about the size of sample, do you take about 1/2 cm? Lets take your pic as an example, where would you take from and which sections would you avoid?


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Re: First time will attempt cloning [Re: Shr00merN00ber]
    #26398917 - 12/24/19 09:45 AM (4 years, 1 month ago)

Quote:

Shr00merN00ber said:
Thanks so much Spunky for your help in all of this. I was wondering about the size of sample, do you take about 1/2 cm? Lets take your pic as an example, where would you take from and which sections would you avoid?




Well I had a TC tell me that 5 o'clock looked good on that plate I just posted. But I'm less than 50 plates deep into this hobby so i cant tell you what i would take from yours...another tc said I could basically take from anywhere on that plate as it all looked pretty good.

I think alot of the time we over think sector selection, I think the main thing is avoiding contaminated parts.

I also have been practicing taking very small pieces like the size of a grain of rice. I even just scraped a bit off of the edge with a scaple and stabbed the middle of the new plate and that worked well...

Thw smaller the better IMO...probably the only time you'll hear that in life lol



Edited by SpunkyMonkey88 (12/24/19 09:50 AM)


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Re: First time will attempt cloning [Re: SpunkyMonkey88]
    #26398921 - 12/24/19 09:49 AM (4 years, 1 month ago)

Quote:

SpunkyMonkey88 said:
Quote:

Shr00merN00ber said:
Thanks so much Spunky for your help in all of this. I was wondering about the size of sample, do you take about 1/2 cm? Lets take your pic as an example, where would you take from and which sections would you avoid?




Well I had a TC tell me that 5 o'clock loomed good on that plate I just posted. But I'm less than 50 plates deep into this hobby so i cant tell you what i would take from yours...another tc said I could basically take from anywhere on that plate as it all looked pretty good.

I think alot of the time we over think sector selection, I think the main thing is avoiding contaminated parts.

I also have been practicing taking very small pieces like the size of a grain of rice. I even just scraped a but off of the edge with a scaple and stabbed the middle of the new plate and that worked well...

Thw smaller the better IMO...probably the only time you'll hear that in life lol





Haha that last part made me laugh LOL. I'll post some pics after pour and some after transfer in a few days. Merry xmas everyone and thanks again for your help!


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Re: First time will attempt cloning [Re: Shr00merN00ber]
    #26398938 - 12/24/19 09:58 AM (4 years, 1 month ago)

No problem man, merry Christmas!


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Re: First time will attempt cloning [Re: SpunkyMonkey88]
    #26399015 - 12/24/19 11:41 AM (4 years, 1 month ago)

Just got done pouring and wrapping with parafilm. Man that parafilm can get annoying with cutting and stretching etc. Here is how it looks. I dont know why I got alot of condensation on some plates (not all), I had the bottle in a water bath at 47C for a while before. Some dishes have these bits in it, all good or  ones with bits looking trash? I seem to find the pieces settled on the bottom.









Cheers!


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InvisibleSpunkyMonkey88
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Re: First time will attempt cloning [Re: Shr00merN00ber]
    #26399138 - 12/24/19 01:12 PM (4 years, 1 month ago)

Quote:

Shr00merN00ber said:
Just got done pouring and wrapping with parafilm. Man that parafilm can get annoying with cutting and stretching etc. Here is how it looks. I dont know why I got alot of condensation on some plates (not all), I had the bottle in a water bath at 47C for a while before. Some dishes have these bits in it, all good or  ones with bits looking trash? I seem to find the pieces settled on the bottom.









Cheers!




Looks good man! And dont worry about the sediment itll still function just the same...

I'm weird about that too so if you want to avoid that next time make twice as much LME as you need and put in a canning jar in the fridge over night, then siphon the top half off with what ever you can (i used a Turkey baster) then run that through a coffee filter, then add your agar for some clear ass agar, If that's what you're looking for...

Anyway that whole process is purely for astectic reasons and you could probably skip either the coffee filter or the s
Mason jar/ siphon step and still get pretty clear agar...

Good work man pouring plates is a huge milestone!


Edited by SpunkyMonkey88 (12/24/19 01:16 PM)


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InvisibleSpunkyMonkey88
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Re: First time will attempt cloning [Re: SpunkyMonkey88]
    #26399142 - 12/24/19 01:14 PM (4 years, 1 month ago)

Also the coffee cup with hot water ontop of your plates trick works like a charm for the condensation :wink:


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OfflineShr00merN00ber
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Re: First time will attempt cloning [Re: SpunkyMonkey88]
    #26404095 - 12/28/19 05:17 AM (4 years, 1 month ago)

Hey guys, hope you are doing well. So, I've just noticed this on one of my plates. It seems like a very small side colony, maybe its contam or a tiny piece of mycelium that fell off during the agar transfer. :crazy:



So my question is, if I want to check the petris in greater detail, do I need to go back to the SAB and take them out of the ziplock bags in there? All dishes are wrapped in parafilm. I dont want to open them, just check each individual dish in detail with a light shining on them. Right now I'm just trying to see them through the outside of the bags, lifting each one and checking.

Thanks!


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OfflinePhycus
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Re: First time will attempt cloning [Re: Shr00merN00ber]
    #26404238 - 12/28/19 08:12 AM (4 years, 1 month ago)

if everything is wrapped in parafilm, you really shouldnt need the ziplocs.


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disclaimer - nothing i post is real. this account is for fictitious purposes and posts should not be taken literally


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Offlinetrubblesome
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Re: First time will attempt cloning [Re: Phycus]
    #26404253 - 12/28/19 08:23 AM (4 years, 1 month ago)

yeah you could play frisbee with one if you wanted as long as it's got something keeping it closed.

you don't have to wrap them until you've put something on it, otherwise you're going to waste a lot of parafilm - once they're set stack them and put the sleeve they came in over the stack. flip them over, fold a paper towel in to a square and put it on top, then tape the sleeve shut. the paper towel will help cut down on condensation. You can stack them upside down before putting the sleeve over the stack so that they're right side up in the bag, but I like taking them out of the bag better when they're in it upside down as it's more difficult to accidentally remove the top of one and potentially contaminate it.


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OfflineShr00merN00ber
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Re: First time will attempt cloning [Re: trubblesome]
    #26404358 - 12/28/19 09:39 AM (4 years, 1 month ago)

Here are some pics from two of my petris. To be honest, I didn't wash my hands and handled them 'carefully' with my bare hands. Here is a plate I feel like could be contaminated to my untrained eyes.

Both the following and the previous plate I posted came from the same cluster tissue sample:

Front



Back



Back zoom




Some info from experienced eyes would be nice.



Here is another petri just for fun from the big clone tissue sample.

Front



Back



Cheers!


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Re: First time will attempt cloning [Re: Shr00merN00ber]
    #26405106 - 12/28/19 05:34 PM (4 years, 1 month ago)

Got a couple of questions I'm wondering about (sorry for the double post). What is the best method for spawning bulk while trying to isolate genetics.. g2g or lc?

What is the general consensus on the best method when taking speed and reliability (less chance of contam) into consideration?


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InvisiblefahtsterM
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Re: First time will attempt cloning [Re: Shr00merN00ber]
    #26405284 - 12/28/19 07:23 PM (4 years, 1 month ago)

G2g will be a bit faster but it’s riskier because you’re opening the jar/bag... LC can be inoculated through a self-healing port. 

Agar to agar transfers are your best bet for isolation.. not sure how lc’s and g2gs are going to be better than one another for isolating.. if you mean which is better for reducing genetics from an MS inoculation, I’d say g2gs since they’re somewhat stationary whereas, lcs are free floating in liquid; you’ll end up with more sub strain colonies.

Since you already have agar going, look into https://www.shroomery.org/forums/showflat.php/Number/24740168/fpart/1/vc/1 josex biopsy method for LCs (sorry if it’s already been said—I’m on the move)

Faht


Edited by fahtster (12/28/19 07:33 PM)


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OfflineShr00merN00ber
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Re: First time will attempt cloning [Re: fahtster]
    #26405614 - 12/29/19 02:27 AM (4 years, 1 month ago)

Awesome, thanks for the info Faht. Do you see any problems with the petris above? In a couple days I'll post a collage with all petris so maybe someone can help me choose the best plate for g2g.


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