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#1 SelfTransformingMachineElf

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Posted 26 April 2021 - 07:48 PM

This will be my last new update thread.  The next thread I start (unless it's a specific question) will just be a running cultivation thread.

 

First - I can't find my digital camera.  The search continues.  Here's to hoping.

 

So, the most imminent update is that I have one half pint Treasure Coast (TC) jar that I will DEC some time this week - probably Friday.  My hope for this jar is a nice isolate, which brings me to the next update:

I'm enjoying some success with my first attempt at agar work!  I thought it would grow a little faster - I've got a tiny bit of Goliath growth and a tiny bit of Mexicana (hereafter Jalisco) growth, but they seem sort of stalled out.  My TC agar didn't take I guess.

 

I did my first set of no-pour transfers last Friday evening.  This was from a very vigorous ATL#7 plate - by far my best grower so far.  I put a little in an LC, a little in RGS qts, and then made 6 transfers to the no-pour plates.  I was hoping to see a little growth in the LC today - no luck.  In fact, all of my LCs suck.  I'm pretty good at following directions - I'm not sure what's going wrong.

 

Everything seems to be growing, if slowly.  I started Ps. cambo - Sandose on Friday evening, and reishi.  I'll be birthing that PF cake, and then continuing to wait for some jars to finish to spawn to bulk.



#2 SelfTransformingMachineElf

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Posted 26 April 2021 - 07:55 PM

To add:  Wow.  I'm reading about Pajarito's experience with the ATL#7 - 16 days from MS to transfer - dang...  I'm not sure how much I'm at, but I'm way behind that.  I've been feeling like things are going slow.  Ah well, I'll feel fine when stuff starts coming in.



#3 SelfTransformingMachineElf

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Posted 27 April 2021 - 04:14 PM

Well - I checked on everything today - more of the same with the LC: nothin'.  My agar transfers are moving very slow - I thought it would be faster!

 

But this check-in is to say that when I checked my ATL PF jars, that they all are producing stones, although they're nowhere near colonized.  Is that normal?  Is that okay?  Is it good?  It seems like fast stone formation would be a desirable trait - at least from a yield standpoint.  Sure do wish I could find that camera.  :(


Edited by SelfTransformingMachineElf, 27 April 2021 - 04:16 PM.


#4 Oldpunk

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Posted 27 April 2021 - 04:41 PM

So...how long has the LC been seeded?

I'm curious to hear what people who know have to say about the stone growth too. I'm still wicked new to all this and I'm really excited to be starting some tampanensis (also a stone grower).

I used some of the tamp spores to make LC almost 2 weeks ago and its just starting to show some tiny specs growing in it. That's the closest thing to an answer I've got, but look forward to seeing others input on your questions.

#5 SelfTransformingMachineElf

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Posted 27 April 2021 - 05:46 PM

April 13th was when I inoculated my first batch - so two weeks today. I was actually thinking it was three weeks - shows my impatience.

 

I grew the stones once before, but it was basically just an inoculation and then I didn't even look at them for three months.



#6 Sidestreet

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Posted 27 April 2021 - 05:47 PM

How exactly are you doing the LCs?



#7 YoshiTrainer

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Posted 27 April 2021 - 06:42 PM

Are you using an SAB or fan while doing your transfers, sterilize between them? Temperature and water content can make a difference in how myc runs your agar. Overly nutritious agar can make for fluffy myc that doesn't want to stretch. Are you using petri dishes or jars for your no pours? If dishes, do you wrap the edges w/parafilm to keep moist? (I'm a jar guy).

I don't do many LCs but liquid agar works great and for another option, Hyphaenation made an awesome thread on milking grains!

https://mycotopia.ne...l/#entry1476763

Good luck!

#8 Oldpunk

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Posted 27 April 2021 - 07:10 PM

April 13th was when I inoculated my first batch - so two weeks today.


So I made mine on 14th I think. And today I had to hold the jar right underneath the grow light and I could barely make out a few tiny specs. And I mean TINY. Maybe your strain is a slower grower?

He he that rhymed

#9 SelfTransformingMachineElf

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Posted 28 April 2021 - 07:42 AM

@Oldpunk - you're a poet and you didn't even realize it!

 

@Yoshi - I'm using a SAB.  My recipe is 10g agar, 10g LME, 1g NY (this time) in 500 mL water.  I would definitely characterize the growth as fluffy, by and large - there are some ropey strands as well - although many of those are on the jar sides.  Since I'm in a SAB and I use profuse isopropyl, I don't use a lamp - I just wipe down my poker thing (technical term).  The first time I tried to get some culture, I realized that I was probably just killing it with the iso, so I started taking bigger chunks to make sure there was some good material that didn't get affected by the iso.  This was my first transfer and second time making an agar batch at all - so definitely still learning.  I'm using PP5 kitchen-style containers and 2 SHIP wide-mouth half pints.  The PP5 containers have a small hole with medical tape.

Is there any way to salvage the fluffy growth?

 

@Sidestreet - the two week old LCs were made as such:

Sugar source was either LME powder OR (real) honey.  I did 2 tsp sugar source into 250 mL warm water - double SHIP lids.  PCed for 30 minutes @ 15 PSI.  The second batch of LC I made was inoculated this past Friday (5 days), and was the same except I used 1.5 tsp sugar source to 250 mL warm water, instead of 2 tsp.


Edited by SelfTransformingMachineElf, 28 April 2021 - 07:49 AM.

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#10 sandman

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Posted 28 April 2021 - 12:17 PM

You can not rely on alcohol to sterilize your tools. Alcohol is only for general cleanliness of the tool, the business end needs to be flame sterilized. You can just have the torch outside your SAB and slowly pass arm thru hole. 

 

Now about that PP5 business...Just called PP! 



#11 YoshiTrainer

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Posted 28 April 2021 - 05:07 PM

If your growth is clean and fluffy, you are fine. If it is fluffy but possibly contaminated, you're still fine, this is where the fun of agar comes in. I started out with the same ratio of LME to agar as you are using and slowly reduced the sugar source over time. My most recent run was 2:1 ratio of agar:DME.

I mix 1/2C agar and 1/4 LME in a tupperware.

For each 1/4 pint jar:
1/4 tsp agar:LME mix
10 ml water

I haven't used nutritional yeast yet but sometimes use charcoal or powdered chicken feed. Ive also soaked sawdust, rice, straw and poo to use the water for making agar.
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#12 Sidestreet

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Posted 28 April 2021 - 05:57 PM

 

Sugar source was either LME powder OR (real) honey.  I did 2 tsp sugar source into 250 mL warm water - double SHIP lids.  PCed for 30 minutes @ 15 PSI.  The second batch of LC I made was inoculated this past Friday (5 days), and was the same except I used 1.5 tsp sugar source to 250 mL warm water, instead of 2 tsp.

 

I'm learning that LME LCs only need a very small amount of LME powder.  I've heard between .2 g per 100 mL and 1 g per mL.  The ones I have now are 1 g per 100 mL and they're still a bit cloudy for my liking, though they are growing.  I think that's too much honey, too.  Maybe just 1 tsp honey per 250 mL water?  Also I would only PC for 20 minutes at the most at 15 PSI with the honey.



#13 SelfTransformingMachineElf

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Posted 28 April 2021 - 09:39 PM

Thanks all.

@sandman - I've always had great success with iso wipe-downs, although this is admittedly my first foray into agar work.  Prior, I was wiping down syringes for LC work, but those syringes were all pre-sterilized and their foil pouches were opened in the pretty sterile SAB.  I actually think I'm going to give hypochlorous acid a try - much nicer smell, doesn't wreck the hands (I don't wear surgical gloves in the SAB, I iso my hands to death), and from some preliminary research, could work better than iso.  AND, then I could even toy around with the idea of putting the lamp IN the SAB because it isn't combustable.  Although then I'm putting a fire source in a cardboard box that has my hands in it - so that may not be great.

 

@Yoshi - I'm not one to be shy of the concept that "less is more".  I'll gladly knock my sugar ratio down.

 

@Sidestreet - I think that a tsp of LME probably weighs somewhere around 4 g - may test that this weekend.  I'll bump down further and see how I do!


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#14 sandman

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Posted 29 April 2021 - 05:17 AM

you can not put a lamp in a sab even with no alcohol because then you don't have still air.


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#15 SelfTransformingMachineElf

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Posted 29 April 2021 - 07:46 AM

good point



#16 SelfTransformingMachineElf

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Posted 29 April 2021 - 09:47 PM

Tomorrow's the day I birth my first cake!  I've got one TC half pint completely finished.  I'm going to DEC it in a small little grow tray with a humidity dome.  Hoping I get a nice fruit to start an isolate.  I imagine I'll always fiddle with looking for "the best" isolate I can find.  Although I say that like I have lots of extra time.

 

I'm trying to remember a few things about DEC:  should I mix coir and verm, or is it straight verm?  Do I moisten and then sterilize, or do I skip the sterilization?  Or do I sterilize dry and then moisten.  I'm leaning toward moisten verm and sterilize right now.  I love a coir&verm casing, but looking through the archives, I never saw any mention of anything other than straight verm.


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#17 Oldpunk

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Posted 29 April 2021 - 10:34 PM

Happy birthing day!

At this point I dont think you need to sterilize. I just rolled mine in verm right from the bag.

I don't know what DEC is. But im new here....and a lil bit high.
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#18 rockyfungus

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Posted 30 April 2021 - 02:26 AM

What's DEC? Is your cake going to bulk, or are you trying to fruit the cake? Coir is a relatively new way of growing mushrooms. We used to believe you needed manure, hay, and nutritious substrates that could lead to contamination.

Technically verm and coir are non-nutritious so you just need to get them to field capacity.  Cakes get dusted in verm and dunked (Haven't done this personally in a decade maybe...). Bulk you shred the cake to coir or coir/verm. 



#19 Sidestreet

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Posted 30 April 2021 - 03:57 AM

DEC is "double-ended casing" for cakes.  I just means putting cakes into the fruiting chamber with field capacity damp vermiculite underneath and on top.  Don't forget to roll the side of the cakes in it too! 

 

I don't know about mixing the verm with coir, never tried it.



#20 rockyfungus

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Posted 30 April 2021 - 07:25 AM

For your DEC no need to sterilize, a colonized cake is really resilient. Just dunk the cake in some clean preferably non-chlorinated water (doesn't seem to mater that much, but why not!) Then roll in your dry verm straight out of bag and set it in your chamber. (forget how often to mist, but I believe you want that humidity high high with cakes)

 

If you are doing DEC no need for coir. CVG is pretty popular (coco,verm, gypsum). I do straight coir most of the time, little bit of verm if I'm struggling to maintain surface conditions during winter.






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