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Offlinesuperwhitedolemite
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Agar Spore Transfer: How am I doing?
    #26427890 - 01/11/20 10:57 AM (4 years, 18 days ago)

My first cloning attempt went horribly wrong and I ran out of fresh ones to try again. So, I grabbed the weakest spore print I had and tried a direct transfer to some of my agar plates, last Saturday.

It's looking pretty good on 2 (or 3?) out of 5 of them, but wanted some feedback from the lot of ya's.

I was super careful...worked in SAB that was vigorously wiped down, in a super clean room, wore gloves, wiped everything down thoroughly with alcohol, etc. I made a small copper loop and flame sterilized between each spore swab (touching the agar to cool, first.)

Still managed to get a significant amount of contamination, but at least two of these plates are looking very strong. Still, I'm surprised at the amount of bacteria here. Seems most likely that the print itself carried some from the cap?

One seems to be thriving so well that it may actually be killing the bacteria? The mycelium is fattening up and the contam seems to be shrinking.

So, the question is; should I attempt to transfer healthy samples to new plates? Is it smarter just to toss the bad ones entirely?

Another question is; how long can I store a fully colonized plate? How do I do this? If even two of these succeed, I'll have way more than I'll know what to do with. I can manage maybe a few tubs in what little spare time I have.

UPDATE 1-19-20:

Wiped out all the crappy pics and got some decent ones. The warm glass of water on the lid to clear the condensation was genius!



Edited by superwhitedolemite (01/19/20 09:18 AM)


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Re: Agar Spore Transfer: How am I doing? [Re: superwhitedolemite]
    #26427895 - 01/11/20 11:01 AM (4 years, 18 days ago)

If those are lit from above, I don't see anything useful here.


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Offlinesuperwhitedolemite
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Re: Agar Spore Transfer: How am I doing? [Re: LadysKnight]
    #26427902 - 01/11/20 11:07 AM (4 years, 18 days ago)

Lit from underneath. That's a ceiling fan/light behind the plates.


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Re: Agar Spore Transfer: How am I doing? [Re: superwhitedolemite]
    #26427904 - 01/11/20 11:09 AM (4 years, 18 days ago)

More difficult to see without light on top.


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Re: Agar Spore Transfer: How am I doing? [Re: superwhitedolemite] * 1
    #26427915 - 01/11/20 11:18 AM (4 years, 18 days ago)

Quote:

superwhitedolemite said:
So, the question is; should I attempt to transfer healthy samples to new plates? Is it smarter just to toss the bad ones entirely?




Thats whats ya do. Taking some normal pics of your petris would help us. We dont want an artful representation of your work. Here's an example of a pic that would be considered helpful....


Gotta be able to see! See?


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Offlinesuperwhitedolemite
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Re: Agar Spore Transfer: How am I doing? [Re: LadysKnight]
    #26427924 - 01/11/20 11:24 AM (4 years, 18 days ago)

Quote:

LadysKnight said:
More difficult to see without light on top.




Fair enough, apologies. I figured it'd be easier to see detail that way but I suppose discoloration would just confuse more than anything.

Here are some w/ no backlight, from above - though this is the underside of the plates. I'm keeping them this way so condensation doesn't drip into the agar, from the lid. I've heard this is smart...seems to make sense for contam prevention.

The two on the left (full pic) are looking great. The three on the right are clearly contaminated - the one w/ the fried-egg-looking contam in the center is the one I asked about maybe pulling through and fending off the bad stuff. Circled the trouble spots in red.



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Re: Agar Spore Transfer: How am I doing? [Re: superwhitedolemite]
    #26427939 - 01/11/20 11:37 AM (4 years, 18 days ago)

Although it would be easier to tell you something if you'd take pics of the top of the plates, with light, it looks like you have some transferrable edges in the first pic. I still have to wonder what the top of the plate looks like.


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Re: Agar Spore Transfer: How am I doing? [Re: superwhitedolemite]
    #26427940 - 01/11/20 11:38 AM (4 years, 18 days ago)

Trash the really shitty ones, and take transfers from those which have mycelium that aren't covered in contamination. I find having to do about 10 transfers from spore until I end up with something that is clean, and grows in an organized and predictable manner.


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Re: Agar Spore Transfer: How am I doing? [Re: WeavieWonder] * 1
    #26427950 - 01/11/20 11:45 AM (4 years, 18 days ago)

Quote:

WeavieWonder said:
Trash the really shitty ones, and take transfers from those which have mycelium that aren't covered in contamination. I find having to do about 10 transfers from spore until I end up with something that is clean, and grows in an organized and predictable manner.




I guess I'm just lucky.. I can usually get some nice looking T3's worthy of grains... I mean they're nowhere near a monoculture (not even sure I know what that is) but I've definitely gotten clean plates from spores 3 transfers in.

I'm new to this so take my opinion with a big grain of salt. But those plates all look contaminated AF to me.

OP how did you inoculate them, apologizes if you said in your OP I'm kinda rushing through the forums


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Re: Agar Spore Transfer: How am I doing? [Re: LadysKnight] * 1
    #26427962 - 01/11/20 11:53 AM (4 years, 18 days ago)

Quote:

LadysKnight said:
Although it would be easier to tell you something if you'd take pics of the top of the plates, with light, it looks like you have some transferrable edges in the first pic. I still have to wonder what the top of the plate looks like.




I agree. Take some photos from above the plates. If they have condensation you can heat up some water in a cup and put it on top of the plate to dissipate the condensation.


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Re: Agar Spore Transfer: How am I doing? [Re: WeavieWonder]
    #26427989 - 01/11/20 12:10 PM (4 years, 18 days ago)

Quote:

WeavieWonder said:
Trash the really shitty ones, and take transfers from those which have mycelium that aren't covered in contamination. I find having to do about 10 transfers from spore until I end up with something that is clean, and grows in an organized and predictable manner.




Yikes! That's fairly time and labor-intensive. This is a fascinating hobby but maybe agar, cloning, etc. is maybe a little too deep for the amount of time I can invest. Perhaps it's straight to syringe with new prints, from now on.

What's the advantage of it growing in an "organized and predictable manner"? I personally don't care if they're pretty or not, just if I'm able to extract samples for inoculating jars.


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Offlinesuperwhitedolemite
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Re: Agar Spore Transfer: How am I doing? [Re: rido]
    #26427991 - 01/11/20 12:12 PM (4 years, 18 days ago)

Quote:

rido said:
Quote:

LadysKnight said:
Although it would be easier to tell you something if you'd take pics of the top of the plates, with light, it looks like you have some transferrable edges in the first pic. I still have to wonder what the top of the plate looks like.




I agree. Take some photos from above the plates. If they have condensation you can heat up some water in a cup and put it on top of the plate to dissipate the condensation.




First I'm hearing of that. Have a quick link for reference? OK, will look at doing this over the weekend and will try to come back w/ better pics.

Thanks!


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Re: Agar Spore Transfer: How am I doing? [Re: superwhitedolemite]
    #26427999 - 01/11/20 12:15 PM (4 years, 18 days ago)

10 transfers is pretty extreme, I think most people drop to grain at t2-t3.

Plus with agar you can just wait til you have a bunch of plates you want to do then take transfers in one session and you’re done for a week or two. I can usually set everything up do 5-10 transfers and wrap the plates in under 30 mins.

The heat on the plate thing works great, sometimes just heating up the room a little will do it too.


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Edited by A.k.a (01/11/20 12:17 PM)


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OfflineWeavieWonder
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Re: Agar Spore Transfer: How am I doing? [Re: superwhitedolemite] * 1
    #26428011 - 01/11/20 12:24 PM (4 years, 18 days ago)

Quote:

superwhitedolemite said:
Quote:

WeavieWonder said:
Trash the really shitty ones, and take transfers from those which have mycelium that aren't covered in contamination. I find having to do about 10 transfers from spore until I end up with something that is clean, and grows in an organized and predictable manner.




Yikes! That's fairly time and labor-intensive. This is a fascinating hobby but maybe agar, cloning, etc. is maybe a little too deep for the amount of time I can invest. Perhaps it's straight to syringe with new prints, from now on.

What's the advantage of it growing in an "organized and predictable manner"? I personally don't care if they're pretty or not, just if I'm able to extract samples for inoculating jars.




Contamination is REALLY good at hiding. If there are a bunch of mycelium strains dog-piled on top of each other, contams can hide in between the layers, making them hard to see.

It doesnt always take ten transfers. Sometimes you get a clean print, and you only gotta do a few transfers. It also helps if you start out with as little spores as possible.

If you're looking for easy, just do the pf tek. It's known for its unpredictability and low yields. Have fun.


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Re: Agar Spore Transfer: How am I doing? [Re: WeavieWonder] * 1
    #26428028 - 01/11/20 12:31 PM (4 years, 18 days ago)

Lol pf tek is the worst. Takes forever, yields shit, and takes way more maintenance than a tub.

I think coir shoeboxes should be the default entry grow. Much simpler than cakes ime, not to mention the much larger yield.


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Offlinesuperwhitedolemite
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Re: Agar Spore Transfer: How am I doing? [Re: A.k.a]
    #26428041 - 01/11/20 12:39 PM (4 years, 18 days ago)

Quote:

A.k.a said:
Lol pf tek is the worst. Takes forever, yields shit, and takes way more maintenance than a tub.

I think coir shoeboxes should be the default entry grow. Much simpler than cakes ime, not to mention the much larger yield.




Yeah, I agree. So far, all I have under my belt is one (seemingly) successful pf tek grow. Did 11 cakes and got around 2 oz, dried. Zero contam of any kind and it was fairly easy...EXCEPT...yes, it was super labor-intensive. Picking a few cakes here-and-there, re-dunking, etc. It was eating up a lot of hours I didn't have.

My thoughts exactly - a tub is just a big cake. I'm only producing for my own use, so I don't need massive yields and giant grows. I bought a bunch of small tubs - larger than shoeboxes but not by much. Pure part-time hobbyist here.

You're not the first to point me toward shoeboxes so if you've got a set of instructions to follow for a simple tek, to point me in the right direction, I'd love to see.

Thanks!


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Re: Agar Spore Transfer: How am I doing? [Re: superwhitedolemite]
    #26428102 - 01/11/20 01:20 PM (4 years, 18 days ago)

Lol yeah I started as a part time hobbyist so I’d have a stash for myself also...three months later I’ve got a whole agar station with hundreds of plates and half gallon mason jars full of mushrooms with stacks of shoeboxes growing.

It’s just what happens.


There’s a few shoebox threads but really you don’t need them. The important thing is grainspawn threads. All a shoebox is is a quart of spawn mixed with a quart or two of coir then you snap the lid on and that’s it.


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Edited by A.k.a (01/11/20 01:20 PM)


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Offlinesuperwhitedolemite
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Re: Agar Spore Transfer: How am I doing? [Re: A.k.a]
    #26428152 - 01/11/20 01:45 PM (4 years, 18 days ago)

Quote:

A.k.a said:
Lol yeah I started as a part time hobbyist so I’d have a stash for myself also...three months later I’ve got a whole agar station with hundreds of plates and half gallon mason jars full of mushrooms with stacks of shoeboxes growing.

It’s just what happens.


There’s a few shoebox threads but really you don’t need them. The important thing is grainspawn threads. All a shoebox is is a quart of spawn mixed with a quart or two of coir then you snap the lid on and that’s it.




Ha! I'm trying to be careful not to get sucked into it like that. I could! But, I just don't have the bandwidth for that right now.

I've watched a couple vids on both and it all seems pretty simple. I thought I'd go for the gusto and try Bod's Unmodified Tek in my "shoeboxes":

https://www.shroomery.org/forums/showflat.php/Number/22337800

Thanks for the info!


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Re: Agar Spore Transfer: How am I doing? [Re: superwhitedolemite] * 1
    #26428381 - 01/11/20 03:36 PM (4 years, 18 days ago)

Quote:

superwhitedolemite said:
Ha! I'm trying to be careful not to get sucked into it like that. I could! But, I just don't have the bandwidth for that right now.

I've watched a couple vids on both and it all seems pretty simple. I thought I'd go for the gusto and try Bod's Unmodified Tek in my "shoeboxes":

https://www.shroomery.org/forums/showflat.php/Number/22337800

Thanks for the info!




I wouldn't recommend that, especially just starting out. If you want to do a shoebox, follow Bod's shoebox tek.


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Re: Agar Spore Transfer: How am I doing? [Re: rido] * 1
    #26428393 - 01/11/20 03:43 PM (4 years, 18 days ago)

Personally I’d say you’d be much better off running multiple shoeboxes instead of a mono. That way all your eggs aren’t in one basket, and you’ll get to see three different grows at once so you could try adding more/less water to one or whatever.

Idk what your goal is but each shoebox could very easily put out two ounces, the worst I’ve ever had was around 18g dry and it was super bad spawn and went moldy at the end of the first flush.

Like I said I started out initially just wanting a stash of shrooms and it sounds so difficult reading about it I started 7 quarts of seeds expecting them to not all make it.

Well they all did fine and all of a sudden I was harvesting a half ounce to an ounce every other day for a couple weeks lol.


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Re: Agar Spore Transfer: How am I doing? [Re: A.k.a]
    #26441418 - 01/19/20 09:45 AM (4 years, 10 days ago)

Man, you're killin' it! I want to have all of these problems. Haha!

I'll go with shoeboxes then - what you're saying makes perfect sense. As I up my skills, maybe then move to a bigger tub.

rido, thanks for the link!

I've updated the OP with much better pictures. Would appreciate feedback from those that recommended I do this. Thanks!

Here are some follow-ups from this morning as well. I'm watching them change by the hour!

I rolled the dice to see what would happen. I carved up the cleanest looking plate - gen 1, no transfer - and threw the colonized agar into a few rye grain jars I prepped the other night. Two days in and it's actually looking pretty solid, no?

Thanks again, all.



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Re: Agar Spore Transfer: How am I doing? [Re: superwhitedolemite] * 1
    #26441432 - 01/19/20 09:55 AM (4 years, 10 days ago)

Thankfully aka hopped in here...

All of those plates look fine but need more transfers. You’re not going to get canopies from any culture like that unless you have extremely optimal surface conditions. Furthermore, don’t waste too much time on MS (multispore cultures), just take a few edges, that look similar, grain them, and then spawn it all andfind a good clone. Clusters are always good clones. Then your working with some predictable and real.


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Re: Agar Spore Transfer: How am I doing? [Re: superwhitedolemite] * 1
    #26441433 - 01/19/20 09:55 AM (4 years, 10 days ago)

just keep a close eye on these. ten transfers is a lot but at least one is recommended. show us what tubs you have and we can help you find a tek that more closely matches them.


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Re: Agar Spore Transfer: How am I doing? [Re: trubblesome]
    #26441447 - 01/19/20 10:02 AM (4 years, 10 days ago)

Quote:

jbgtaa said:
Thankfully aka hopped in here...

All of those plates look fine but need more transfers. You’re not going to get canopies from any culture like that unless you have extremely optimal surface conditions. Furthermore, don’t waste too much time on MS (multispore cultures), just take a few edges, that look similar, grain them, and then spawn it all andfind a good clone. Clusters are always good clones. Then your working with some predictable and real.




Interesting. So, is it the concentration of the mycelium, or the robustness/health, that makes it more viable on each transfer? I noticed the first transfers I did are much denser and growing in a radial pattern.

Like I said, I just whipped up 3 jars and threw some in there, just to see what would happen. I'll stick these in 3 small tubs when they're ready and see how it goes. Strictly as an experiment to satisfy my curiosity.

I often like to learn things the hard way as well. Life-long affliction.

Quote:

trubblesome said:
just keep a close eye on these. ten transfers is a lot but at least one is recommended. show us what tubs you have and we can help you find a tek that more closely matches them.




Fair enough. No tubs yet, this will be my first run. Will definitely throw a thread out there once they're started. All I've succeeded in doing to date, is 11 cakes a la PF Tek. That's where all the spores for this new round of stuff came from.


Edited by superwhitedolemite (01/19/20 10:03 AM)


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Re: Agar Spore Transfer: How am I doing? [Re: superwhitedolemite] * 1
    #26441452 - 01/19/20 10:05 AM (4 years, 10 days ago)

I really can’t speak on isolating/culturing multispore cultures. I find it to be a waste of time and the returns are usually diminishing considering you have no clue what you’re working with.

When you take a clone, you’ll see what I’m talking about. Similar rhizomorphs, radial patterning, less “piled” mycelium. Check my gallery there’s some solid GT clone cultures, and some not so solid, but youll see how it differs from MS.


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Re: Agar Spore Transfer: How am I doing? [Re: superwhitedolemite] * 1
    #26441459 - 01/19/20 10:09 AM (4 years, 10 days ago)

Quote:

superwhitedolemite said:


My thoughts exactly - a tub is just a big cake. I'm only producing for my own use, so I don't need massive yields and giant grows. I bought a bunch of small tubs - larger than shoeboxes but not by much. Pure part-time hobbyist here.





these tubs

you should keep it all in one thread (this one) instead of starting a new one. so many people reference previous threads theyve made about their projects and assume everyone knows everything theyve done up to that point in their grow and can tell them exactly why this or that is happening. keeping it in one thread will make it easier for people to help pinpoint problems down the road


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Re: Agar Spore Transfer: How am I doing? [Re: jbgtaa]
    #26441472 - 01/19/20 10:17 AM (4 years, 10 days ago)

Quote:

jbgtaa said:
I really can’t speak on isolating/culturing multispore cultures. I find it to be a waste of time and the returns are usually diminishing considering you have no clue what you’re working with.

When you take a clone, you’ll see what I’m talking about. Similar rhizomorphs, radial patterning, less “piled” mycelium. Check my gallery there’s some solid GT clone cultures, and some not so solid, but youll see how it differs from MS.




Sorry, relatively new to all the terminology. That is to say; multispore culture is the result of transferring spores to agar?

I've gotten a lot of comments about how ugly my plates are, so I'm trying to ascertain if that's just preference or if the end result is really radically different. I'm very new, so I'm at the stage where if I get anything to grow and it's not contaminated, I'm super stoked about it. Haha.

Will check out your stuff. I've made one cloning attempt so far that went hilariously wrong - 100% bacterial shitstorm. The fatal mistake was cutting into the mushroom and using the same blade to immediately transfer the sample. I'm definitely going to give it another try on the next flush.


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Re: Agar Spore Transfer: How am I doing? [Re: superwhitedolemite] * 1
    #26441481 - 01/19/20 10:28 AM (4 years, 10 days ago)

I had trouble with bacteria my first few clones also.

Gotta glove up then rip the stem in half in the SAB and make sure the blade doesn’t touch the outside of the stem.

My method at this point is to cut a square shape pretty deep in the center tissue then cut the square in half diagonally downward so a nice little triangle shape comes loose.

I also accidentally discovered the tiniest piece of clone tissue will grow fine. I had a little speck fall off onto a plate and it grew out  same as the actual transfer.


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Re: Agar Spore Transfer: How am I doing? [Re: A.k.a]
    #26441488 - 01/19/20 10:36 AM (4 years, 10 days ago)

Quote:

A.k.a said:
I had trouble with bacteria my first few clones also.

Gotta glove up then rip the stem in half in the SAB and make sure the blade doesn’t touch the outside of the stem.

My method at this point is to cut a square shape pretty deep in the center tissue then cut the square in half diagonally downward so a nice little triangle shape comes loose.

I also accidentally discovered the tiniest piece of clone tissue will grow fine. I had a little speck fall off onto a plate and it grew out  same as the actual transfer.




Noted, thanks! Yeah, we were laughing about that. All that prep, cleaning, etc. and my fatal flaw was cutting into the stem, then cutting the sample out. Everything I did from that moment forward was a waste of time and money. Lol!

Admittedly, I got a little cocky after zero contam on my first attempt with PF, and assumed I was invincible. I took nearly zero precautions on that grow and didn't even have a SAB. I got the impression at that point, that this stuff would be easy. :tongue:


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Re: Agar Spore Transfer: How am I doing? [Re: superwhitedolemite]
    #26441519 - 01/19/20 11:07 AM (4 years, 10 days ago)

i mean, you can use the same blade, just flame sterilize it


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Re: Agar Spore Transfer: How am I doing? [Re: trubblesome]
    #26442657 - 01/20/20 05:56 AM (4 years, 9 days ago)

Right, yeah. That's the one thing I didn't do. Makes sense but the video I followed didn't, so I thought nothing of it.


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Re: Agar Spore Transfer: How am I doing? [Re: superwhitedolemite] * 1
    #26442956 - 01/20/20 10:52 AM (4 years, 9 days ago)

Quote:

superwhitedolemite said:
Right, yeah. That's the one thing I didn't do. Makes sense but the video I followed didn't, so I thought nothing of it.




Also, like aka said, tear it open, don't cut into it. When you cut in from the outside you are pushing contams into the inside.


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Re: Agar Spore Transfer: How am I doing? [Re: LadysKnight]
    #26449897 - 01/24/20 12:21 PM (4 years, 5 days ago)

Right, gotcha. I suppose it'd be just as easy to push contams into the flesh of the stem by ripping it open, without being super careful?

Excited to try cloning again now that I've had more success w/ agar plates. I'll study up before the next attempt. Thanks!


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Re: Agar Spore Transfer: How am I doing? [Re: superwhitedolemite] * 1
    #26449921 - 01/24/20 12:30 PM (4 years, 5 days ago)

As long as you tear it it should be clean. Cutting it you push the blade through the dirty skin and slide it through the clean stuff, which could spread bacteria kind of like butter onto bread.

I pinch the base of the stem to crack it then pull it apart like string cheese, so I’m only ever touching the outside of the base of the stem, then take the sample from the middle/top part. The thicker they are the easier it is to cut a clean piece out.


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Edited by A.k.a (01/24/20 12:32 PM)


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Re: Agar Spore Transfer: How am I doing? [Re: A.k.a]
    #26449950 - 01/24/20 12:43 PM (4 years, 5 days ago)

Round of pics from yesterday. Wish I had time to get more today since everything is so much further along! Bumped the temp in my room the last two days and man, what a difference!

I circled some potential trouble spots. Newer plates (round 2 transfer) look much better so may just ditch the first round. Maybe that's not mold but why risk it, right?

Jars are going nuts. I think I'm maybe 2-3 days away from spawning to tubs. Shook em up once already and they might need it again today.

Some questions I asked initially but probably got lost in the shuffle:

What do I do with plates that I can't use right now? I have more healthy agar plates than I'm going to be able to utilize. Can they be safely refrigerated? Will the mycelium die? If they can be stored, for how long?



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Re: Agar Spore Transfer: How am I doing? [Re: superwhitedolemite]
    #26450024 - 01/24/20 01:24 PM (4 years, 5 days ago)

Are those the jars you’re talking about?

You can store plates at room temp for months, but I don’t think you’ll get much use out of those. Did you pour that agar or is it premade?

None of the plates look normal but it kind of seems like something going on with the agar.


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Edited by A.k.a (01/24/20 01:27 PM)


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Re: Agar Spore Transfer: How am I doing? [Re: A.k.a]
    #26450054 - 01/24/20 01:41 PM (4 years, 5 days ago)

I made the agar - was my first attempt. Yes, these jars are also my first attempt at those.

I mentioned earlier on that I had transferred to these jars directly from a first-gen plate, no transfers. They seem to be doing well! If they don't, no worries, it was only an experiment.


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Re: Agar Spore Transfer: How am I doing? [Re: superwhitedolemite] * 1
    #26450119 - 01/24/20 02:11 PM (4 years, 5 days ago)

Right on.

Well unless they were shaken within a day or so of the pictures they still have a lot of colonizing to do, gotta be solid white.

What was your agar recipe? I’ve been making very soft plates lately and it seems to be affecting the growth now, I’m getting a lot of clear growth at the edges kind of like the myc is growing in the agar rather than on it.

Might be something similar going on with yours.


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Re: Agar Spore Transfer: How am I doing? [Re: A.k.a]
    #26450411 - 01/24/20 05:42 PM (4 years, 5 days ago)

Oh, for sure. These pics are from yesterday. As of today, about 50% of the rye is visibly covered again. It seems to really be picking up all of a sudden.

It’s definitely weird how the myc seems to dig into the agar, then sprout up and move outward again. I hadn’t even considered it might be the recipe. I followed this one to the letter:



What’s your preferred recipe and approach?


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