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OfflineGrambo
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stretching my inoculant
    #24007783 - 01/13/17 04:59 PM (7 years, 1 month ago)

Hello. I spent the summer farting around with PF tek and several "damion 50/50" tubs. I have experimented with liquid and crystal psilocybe and learned a great deal. When I purchased my syringes I took 1 and injected it into a 1/2 pint jar half filled with sterilized water equipped with a SHIP. My theory was to create extra inoculant as the sample was a black blob the size of a pencil eraser. 6 months later I am looking at this giant leech looking blob swimming around in the jar.

I wasn't aware spores multiplied. This happened with only the 1 out of 3 jars that I made so I am not sure what is happening. I only know that this particular jar was an untouched vendors syringe and has inoculated several grows. I'm just curious if anyone can shine light on this. Am I just imagining it is bigger?

I only use boiling water in the syringe once it has cooled or air drawn through a flame when extracting a sample to keep a positive pressure in the vessel and prevent contamination. I always inject through a 70% alcohol swab as well so it's not a blob of bacteria, it is healthy and successful.

Anyway, just wondering... this B+ sample grew me 10 specimens between 3 to 5.5 grams.

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InvisibleAdden
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Re: stretching my inoculant [Re: Grambo] * 1
    #24007809 - 01/13/17 05:09 PM (7 years, 1 month ago)

Spores don't multiply.

You can fill a syringe 75% of the way with water and PC it. If there's air it won't pop.

No need to ISO needles just flame them.

If you're gonna stretch it, honestly just clean it up and start new. Who knows what's in there. You can use the rest and pump out a few cakes or something while you learn agar. The last thing you want is some latent bacteria growing in there and have to throw away tons of spawn.

Liquid inoculant only takes a few days, a half pint jar filled 50% with water, drop agar in it and shake shake shake over a day or two and you're good to knock up whatever.

I'd say learn agar and put some of your LC on it to make sure it's clean and if it is then knock up a ton of grain jars or bags.

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InvisiblebodhisattaMDiscordReddit
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Re: stretching my inoculant [Re: Adden] * 1
    #24007898 - 01/13/17 05:51 PM (7 years, 1 month ago)

Damion5050's tek has absolutely nothing to do with 50/50
So you did those tubs wrong or you didn't really do them

Edited by bodhisatta (01/13/17 06:01 PM)

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OfflineGrambo
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Re: stretching my inoculant [Re: bodhisatta]
    #24007969 - 01/13/17 06:27 PM (7 years, 1 month ago)

Nobody said anything about a half and half mix buddy... I used the tek, sorry if the backslash confused you, but that wasn't the topic either.




This was one of 6 tubs I did this summer... am I a liar?

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OfflineGrambo
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Re: stretching my inoculant [Re: Grambo]
    #24007989 - 01/13/17 06:34 PM (7 years, 1 month ago)

As far as my question thank you for the answer ADDEN.

Here is an excerpt from the "agar and culture storage" from the above menu-

Home | Mushroom Info | Growing Mushrooms | General Cultivation | Agar and Culture Storage | Resurrecting a Better Method for Long-Term Storage of Mushroom Cultures

Resurrecting a Better Method for Long-Term Storage of Mushroom Cultures

Distilled Water:

I came upon an article written by Michael D.Graham,a microbiologist at ATCC (1) that described the storage of yeast in sterile distilled water. What a brilliant idea! If that method stores yeast, It should work well on gourmet mushroom cultures, too. It's easy to do, very space efficient, allows the cultures to be stored at room temperature, and maintains thier viability for years. I contacted the author, he indicated that edible fungi stores even better than yeast, and you can store the spores as well as the hyphae often for decades!

Rating: 10 bells.

Sterile water was first used to preserve cultures by Castellani in 1939 (2). Since then, many scientists have used this method; McGinnis et al. in 1974 (3) and Odds in 1991 (4) reported that they were able to maintain viable cultures for more than three years, without degredation.

This technique satisfies many different interests: Castellani's pathogenic fungi, Odds interest was in pathogenic yeast, and McGinnis' interest was in a wide range of fungi, yeast, and bacteria. The distilled water preserved all of them.



I have had no trouble with my jar yet and if this one fails I will update the post and accept that I was wrong.

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InvisibleAdden
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Re: stretching my inoculant [Re: Grambo]
    #24008072 - 01/13/17 07:09 PM (7 years, 1 month ago)

People use slants to preserve genetics. I don't know how to do them so I just do no pour agar with micropore plastic lids and set them at an angle in my PC and throw them in the fridge.

Agar is worth learning.. if anything, it's worth it for making slants and testing how clean your inoculant is.

Then you can snag genetics off large fruits or potent fruits or clusters of fruits, and repeat that process until you have full canopies of beastly, potent fruits.

Sometimes genetics mess up. So it's always good to have backups of backups. People keep master grain jars for this reason, too.

I hope some of this makes sense to you.

Basically you've got to "build" the kind of mushrooms you want and test genetics occasionally. Pick and choose aggressive mycelium of the best of the best. Eventually you can turn clones into an isolate.. then you just keep making those fruits over and over again. Very much summarized, but this should be every long term bulk grower's goal. If you stop growing for some reason you can just jump right back into it with a piece of myc off a slant.

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OfflineGrambo
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Re: stretching my inoculant [Re: Adden]
    #24008106 - 01/13/17 07:26 PM (7 years, 1 month ago)

I have done a bit of reading on agar but it seems a bit scary to get going. I bought a few packages of agar from an oriental grocer in Calgary, I just can't find petri dishes and I'm thinking of using my 1/2 pint jars if need be.

I'm aware of the cloning concept and I am interested, just trying to figure it all out. Like... how do I determine a good candidate for potency when I would need to ingest it first? In a multispore grow each specimen could be different, other than that it seems like trial and error. It just seems intimidating and I wish there was a good read to get me motivated.

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InvisibleAdden
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Re: stretching my inoculant [Re: Grambo]
    #24008186 - 01/13/17 08:06 PM (7 years, 1 month ago)

Well, you've used a PC before so I assume you've got one.

"No pour" agar, aka "Pasty Plates" are easy.

Bring 1.5 cups of water to a hard boil, add food coloring, add Karo, whisk a pinch of potato flakes in, slowly add 7g of agar.. like tap the edge of a cup or something to get a light dusting. I turn the boil down once the dextrose and potato are in. Sometimes I add a quarter cup of extra water if I see it steaming off.

So now your agar is done. Take some Glad mini round pp5 (the 5 is the recycling code on the bottom). Put a 1/4" hole in the lid. Cover this with 2 layers of micropore tape. I like to have these out and lined up. I also use the pint pp5 ones because they're more like a dish.

So pour about 1-3cm of agar.. you really don't need a lot.. and fill them as evenly as you can. Let them steam off a little but not too long. Put the lids on. Take a folded paper towel and cover your micropore tape. Wrap in foil. PC at 15psi for 45 minutes. Let sit overnight (or a couple hours it doesn't retain heat).

Load them into your Still Air Box (SAB) and let the tape dry. If you have other projects going, do your SAB work in sessions. Either streak plates with sterile swabs from prints or get a small exacto knife set.

This is when you can add spores or tissue to your agar and begin your culture work. Try to keep a few irons in the fire so if it all goes to hell, at least you have grain jars or bags or PF cakes or something going.

It is easier than making jello, and all you do is PC it.

Normally people don't spoon feed information, it's a good way to get flamed or trolled, use the search engine and toggle "earlier than 5 years" for stuff you're doing.

There's an ongoing Q&A thread in the AMU forum.

A.M.U Cultivation Q&A


Here's some stuff I had hanging around that might help you. Not sure if you go picking for wilds but there's some info for that too.

Easy Guide to Describing your Mushroom Specimens * Stipe Texture Variations and Descriptors * Cap and Gill Morphology Reference Sheet * Active Mushroom Species by State * Agar Quicklinks * Frank's 12 step spore-to-bulk * When To Harvest your Mushrooms * Blenderless Liquid Inoculant * Bacteria Morphology Cheat Sheet (agar)

Here's a 13 step cheat sheet on making BRF cakes if you want to get some of those going. You can use as little as half a cc of the LC you have to get them going. They'll finish and fruit fast so you'll have some fresh genes to work with. That's as easy as taking a big fruit or cluster of fruits and taking tissue and moving it to agar. You can start playing around and narrowing down the genetics you want to see in your tubs.

Here's some stuff that may help. Let me know if you need anything else.. I toggled this thread as a favorite so I'll be notified when it's posted in.

As below.. if anything, get the SAB (still air box) constructed if you don't have a flowhood. You'll be using it for the rest of your myco days.

Quote:

Read them in order, invest some time in soaking up all this and you will never regret it:

Why you should start with agar (take a look at the links in the OP as well)

Pasty's plates

Videos on how to work with Pasty's plates and sterile technique

The Tiger Drop - Grow From Your Own Prints With No Contamination - Tek for n00bs

How to make a soft agar mix to germinate spores and to make a blenderless LI

Blenderless LI

Agar Portal

Improve your sterile technique! (a comprehensive guide to agar)

And also very important, watch all of RogerRabbit's videos on agar and sterile technique, they're badass (included in the link "Agar Portal").

Bonus
SAB



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InvisibleAdden
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Re: stretching my inoculant [Re: Grambo]
    #24008197 - 01/13/17 08:11 PM (7 years, 1 month ago)

Quote:

Grambo said:
I'm aware of the cloning concept and I am interested, just trying to figure it all out. Like... how do I determine a good candidate for potency when I would need to ingest it first? In a multispore grow each specimen could be different, other than that it seems like trial and error. It just seems intimidating and I wish there was a good read to get me motivated.




Yes.. it's why I always suggest people keep their SGFC's. Knock up fast with LI/LC, test potency and growth etc and if they're good then grab the best of the best and repeat.. you can run tubs side by side.

Multiple Spores inoculation, yes, it could be anything. Trillions upon trillions of genetic variations. You whittle it down to a couple billion or so by doing agar work. It's the foundation to getting started.

Here's a good book. The Fifth Kingdom. First 5 chapters are free. Everything else you need to know is here on the boards.

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Offlinetump
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Re: stretching my inoculant [Re: Adden]
    #24008210 - 01/13/17 08:20 PM (7 years, 1 month ago)

Op to answer your actual question spores don't triple or grow like that in water i did a similar thing with my pink buffalo grow. After time more spores get in form your filter. My first bacth with one syringe was 95% perfect . a month later i had 100% black mold fail rate form same jar. Just remembet when divideing to use it all at once

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InvisibleAndyHinton


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Re: stretching my inoculant [Re: Grambo]
    #24008301 - 01/13/17 09:24 PM (7 years, 1 month ago)

Update: Reached 50 posts.


--------------------

Edited by AndyHinton (01/26/17 09:38 AM)

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OfflineMysticMoteToter
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Re: stretching my inoculant [Re: AndyHinton]
    #24008319 - 01/13/17 09:37 PM (7 years, 1 month ago)

Quote:

AndyHinton said:
I'm glad you mentioned distilled water and surprised more people don't use it. When you wash 100 mL of grains in it, you get a high quality universal inoculum.



honestly ime i've gotten worse contams using distilled than slightly chlorinated (tap water)


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OfflineGrambo
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Re: stretching my inoculant [Re: MysticMoteToter]
    #24008527 - 01/14/17 12:00 AM (7 years, 1 month ago)

Thanks for the material Adden, I had no idea the search engine had those features so I searched how to use it... thank you, I feel a little stupid now.

I appreciate the advice and the links, you're a good person.

I still have B+, Golden Teacher and Treasure Coast syringes unused in the fridge as well. I just got carried away with trying to stretch my supply. I need to admit I mixed up 2 of the jars, the reason the blob was larger is because it was a different jar. This jar was a suspended mushroom cap over sterilized water print that must have rehydrated somewhat over time in the fridge.
Whatever, I noc'd 6 quarts and 3 1/2 pints to stall in the fridge if they work. In the meantime I am going to crack a beer and watch some tv, tomorrow I will come back for some more searching and reading. I will PM you if I have any stupid questions that I can't find now that I can UTSF.

Thanks again guys.

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OfflineGrambo
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Re: stretching my inoculant [Re: Grambo]
    #24050222 - 01/29/17 08:50 PM (7 years, 1 month ago)

I said I would report back if I failed so here it is...
The grain jars failed with a white dusty almost oxidized look, mostly at the injection point, but everywhere else too. No picture of those, sorry. I left them a bit and the first thing I saw was grey fuzz... the fck outta here.

Here's what I got



I made some PF cakes from ground millet/rye/brown rice and used another syringe of that culture. The black clots can be seen stuck to the container emanating the grey fuzz.

The vendor syringes unused, and 2 with the B+ I made.



I remember the B+ I made was like cracked mud dust floating on the water surface and was hard to mix, kinda stayed flaky. Well I guess this wasn't all that bad... I still have viable spores and I am wondering if I can still animate what may be in the TC agar I made. It became a bit of a goo so I stopped moving it and it has gelled... don't know what to do with it. Still got the syringe, might be a tiny drip left in there...

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