42 replies to this topic

[Jrotten]

Agar to LC. I cut the stipe from the sub with flamed scissors, then the cap from the stipe, again with flamed scissors. I put the stipe aside in the hood while I set the caps up for printing and then I tear the stipe in two bottom to top lengthwise with gloved/alcohol wiped hands. I then use a flamed hobby knife to remove 2 fibers from the inside of the stipe and drop them on a single plate. As soon as I see any clean growth I take the smallest possible sections and transfer them. Eventually if I can get clean plates I will take a single wedge, and drop it into an LC. I've been using corn syrup for it's clarity, but I prefer diluted and filtered grain water for the LC itself. I rotate them on my stir plate. I am currently experimenting with very low nutrient LC of 1% instead of 4%. All I can say is that I don't have as thick of cloud of mycelium in the jars, but my hope is that they will exhaust the sugar supply more readily. The simple sugars seem to make things more prone to contamination down the road.

The original plating of clones and spores was done 5 days ago, the first transfers were done 2 days later, they aren’t quite clean, but if it’s going to work one section will grow away from the bacteria. I might even make a transfer to antibiotic agar.

It should be noted I’m much more ambitious than successful. At the moment I feel overwhelmed and I’ll pull this off as well as I’d hoped.

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Edited by Jrotten, 10 January 2021 - 02:57 PM.

[cujoloki]

Thanks Jrotten. Thats a great explanation. Couldnt be more concise. Im sure you will pull this off splendidly. 

[Jrotten]

A picture of what my FC looks like right now. I have absolutely no clue if it can handle this or if I’m treating all these species fairly. I’ve harvested one tray of pans and one small tray of cubes, but now everything is officially tops off and I’m trying to fruit them all. It smells very mushy when opened!

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[YoshiTrainer]

Hell yeah JR, looks great to me!

[Jrotten]

Pan Jambo pins seem to be doing well. They could probably use some more heat, but that’s hard to maintain right now. 72-75F is what I’m working with it seems.

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[rockyfungus]

So now I know. Single wedges shaken really well are much less likely to contaminate and there’s no real reason to ware wedges. Now I have to increase my timetable. I think G2G will be needed just to buy some time.

Sorry to go back in time a bit. What's your agar recipe? I keep decreasing the grams of agar to nutrients and get wedges that almost liquefy when I shake. Also allows for a very sticky agar for easy transfers.

6-7g agar, 10 g nutrients, 500 ML H2O

[jrh]

I've started reducing the amount of agar as well because I was getting tired of transfers not transferring. Looks like I can drop some more based on your numbers.

[Jrotten]

I use 18g of sugars either MEA, corn syrup, or a combination and 18g agar to 750ml. I vary the sugars depending on the plan... if I’m doing LC’s I usually just use corn syrup in both. If I have issues like the subs I recently tried I will do full MEA. I don’t like my agar too soft because I incubate everything upside down. I’ve had my culture wind up upside down in the lid!

[jrh]

LOL I've had my transfers upside down in the lid. That's why I want a softer agar. :)

[rockyfungus]

Agree with JRH, softer agar so inverting doesn't lose my transfer.

[Jrotten]

I have had that happen a time or two, but I just leave larger transfers right side up for a few hours. How do you put spores to soft agar? When I use soft agar it just gums up my swab and I get a mess.

[jrh]

Fahtster recommended that I make a slice in the agar, and rub the inoculation loop through there. For a swab, use sterile scissors to take a tiny snip of cotton and push it in there.

[Jrotten]

Busy day... harvested the Jambo tray but left a lot of young mushrooms. Pins coming up everywhere still so I soaked it and hope they don’t abort. Took clone samples but I was rough with them, the Jambos were small and dense and I need a new blade! I also swabbed direct from gill to agar in the hood. Trying to take prints. I’m still new to print making. Made T2 on the mystery pan I realized was labeled “pan cyan west Texas” which makes no sense to me, it it wa a large fruit. I’d love a canopy of them.

Also got my first ever Mexicana pins var Chicon Nindo. So now I need to pour more plates to take more clones! Pan tray seems to actually be continuing to pin and mature. I soaked them for two hours. That’s 4 species so far

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[Jrotten]

PIIIINNNNNS!!!!!!

I got pins coming up all over the Chichon Nindo tub and the second Pan Cambo Var Jambo trays kicked up it’s first pin, and the AVC cubensis micro tray put up the first pins of it’s second flush which I need to print.

The only real concern I have is I expected the Tampanesis to fruit first and I’ll never really know if the semperviva are going to pin or not until sometime next year!

[Jrotten]

Lost a tub of semperviva to trich.  Got it before it went green so hopefully that helps.  I also drenched/dunked some things over night. 

The one Jambo tray just keeps producing without every "flushing." I'm getting 1-2g dry every day off that little 4 cup tray which is cool, but harvesting around pins every day gets old.  The other Jambo tray has a "single double meaning" two stipes and one cap.   It's very small and has stayed that way so who knows what's going to happen there. 

The one tray of Mexicana Chichon Nindo keeps maturing pins, but I'm not sure what to make of those.  They seem so tiny. 

I think the Tampanesis have just decided to make stones so I'm not sure what to do with that.

I have my ACV clones just sitting in jars waiting to be spawned for weeks.  I need to get them fruited and get prints.

I also have 3 LC's for a Key Island pan cyan clone, a Semperviva clone that has at least grown out clean, and a pan TX that looks like the healthiest of all, but what do I know.

And finally I have way too many pans going on agar!  I have Pan Jambos germinating plus some clone attempts, I think I'll get at least one, Pan TX on T2, as well as Pan TX T2 clones.

[Jrotten]

Two stripes, one cap. Not tiny anymore. Pan Cambo.

And the Mexicana Chichon Nindo maturing right along.

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[Jrotten]

Rushing today:

Made sub, pasteurizing now.
Pan Jambo tray is pinning flushing again.
Mexicana Chicon Nindo is maturing beautifully and the second tub started pinning
AVC cubes are doing their thing

Put Allenii, Gym Purp, and Subs from spore as well as Zapo and Caeroluscens from wedges
Had a lot of bacteria on my pan Jambo spore plates as well as the clones, but managed to transfer one beautiful colony from one of the germ plates and I made three transfers of the clone, I think one of them will take.

Tomorrow I will spawn 3 AVC clones and 3 trays of Chicon Nindo with the same culture taken from 3 different jars. But will be pretty light spawn rate and I added wheat bran to the compost/coir sub I used last time. Hopefully it doesn’t cause issues.

The Chichon Nindo are such photogenic little soldiers :)

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Edited by Jrotten, 19 January 2021 - 01:06 AM.

[rockyfungus]

Staying busy! I had to cut back on my projects as they were stressing me out.
 

 

I have had that happen a time or two, but I just leave larger transfers right side up for a few hours. How do you put spores to soft agar? When I use soft agar it just gums up my swab and I get a mess.

Have to be very delicate with the swab, I tend to roll it gently on the surface. It really doesn't seem to squish the agar unless I use excessive force
But my preferred way is going to be print->agar, not a huge fan of swabs.

[Jrotten]

Well I have no intention of growing fewer species for a minute, or trying. I have thing that I really feel I should be pulling prints from if possible, but I have decided to try and slow incubation times down. It’s going to take discipline and dedication, but I am incubating my trays and tubs at 70F if they’ll colonize. Pan Jambo, AVC cubes, and Chichon Nindo. I just don’t have the room to do something else honestly because my FC is full. My second FC is full. My incubator is full. My incubation drawer is full.

On the one hand I’m streamlining my processes, like finding a sub for everything except pans and wood lovers, which I’m considering simple modifications to for those cases. On the other if I forget to label ANYTHING or worse, label the lid and not the tray... it all becomes overwhelming quickly. I really want a grow closest with shelves and yes I know god to buil it without damaging my house... And I sure do want to!

It’s about time to harvest these MCN.

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Edited by Jrotten, 19 January 2021 - 10:29 PM.

[Jrotten]

@rocky, I use swabs even on prints. It’s sometimes great and sometimes not. I have failed miserably making myself a loop. Repeatedly.