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OfflineBlue Helix
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Constant-stir LC showing expotential growth in first 3.5 days. (pics)
    #4340904 - 06/27/05 01:11 AM (11 years, 10 months ago)

This is my first LC. It was made of 1% light malt and 1% dextrose. It was started with 1cc of an old LC that had been in storage. It was stirred constantly and kept about 83F. I was surprised by the rapid growth.

3.5 days after inoculation (and ready to be used)


I believe this to be actual mycelium, not mold, and as you can see contant stirring with proper incubation temperatures generates a very dense culture very quickly. It this does turn out to be mold, I'll post back to this thread in a few days.

Assuming this is cube mycelium, one can easily generate a completed culture in 4 days from a single CC of live culture. It would probably take one more day for spores. The rapid development is, I believe, due to the constant stirring, and I think it is well worth the investment in a stirrer (about $30 on Ebay).


Edited by Blue Helix (10/06/05 03:07 AM)


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Invisibleohmatic
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Re: Constant-stir LC showing expotential growth in first 3.5 days. (pics) [Re: Blue Helix]
    #4341960 - 06/27/05 06:48 AM (11 years, 10 months ago)

it looks like very nice flakey mycelium indeed.
i am somehow astonished that it developed in a constantly
stirred medium though, all the cultivators i know using liquid usually
stirr just every two days or so.
peace ohm :mushroom2:


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OfflineCLUTCH
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Re: Constant-stir LC showing expotential growth in first 3.5 days. (pics) [Re: Blue Helix]
    #4341982 - 06/27/05 07:06 AM (11 years, 10 months ago)

Impressive.

Also, I believe that 1% is a weak solution. Wouldn't 4% light it up even quicker or more?


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Offlinexburn
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Re: Constant-stir LC showing expotential growth in first 3.5 days. (pics) [Re: CLUTCH]
    #4342054 - 06/27/05 07:34 AM (11 years, 10 months ago)

its really a 2% solution 1+1=2


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Re: Constant-stir LC showing expotential growth in first 3.5 days. (pics) [Re: Blue Helix]
    #4342166 - 06/27/05 09:29 AM (11 years, 10 months ago)

Can we see a picture of it, with the stir plate off & the myc settled down?


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Offlinedebianlinux
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Re: Constant-stir LC showing expotential growth in first 3.5 days. (pics) [Re: Blue Helix]
    #4342394 - 06/27/05 11:29 AM (11 years, 10 months ago)

err, i get the same degree of propagation just shaking the hell out of the jar once a day. My LC's are 100% ready for use in exactly 4 days after inoculation.

i, too, would like to see the settled mass to determine if the constant stirring actually resulted in more mass in the same time period.


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OfflineBlue Helix
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Re: Constant-stir LC showing expotential growth in first 3.5 days. (pics) [Re: agar]
    #4342403 - 06/27/05 11:33 AM (11 years, 10 months ago)

agar, I would love to show you that solution settled, but I just used it! When I allowed it to settle for about 5 minutes, I estimate about half the jar looked to be a mycelium cloud, like the "done" pictures I've seen. The other half was more clear looking.

And I made a mistake: this is a 4% solution, 2% powdered dextrose and 2% extra light malt. I put the sugars in a double coffee filter and ran hot distilled water through the coffee machine (thank's Hippiechick for that idea). I actually heated the water in the microwave prior to putting it through the coffee machine to try to make it hotter.

With this particular extra light malt--which the beer guy was all excited about as the best light malt in the world or some such--I did not see any residue left in the filters when I was done, but it might have just been too light to see. That gave me a reasonably filtered 500 ml solution which I pressure cooked for 20 minutes (as an aside it was not quite as crystal clear filtered as I've seen on here from agar and the like).

I am not sure why the liquid culture developed so quickly. I was astonishing and did not expect it since I have never done a live culture before. It developed so fast that I am still fearful that it might not be cubensis mycelium, but I have no indication otherwise at this point. If it turns out to be green mold or something, I'll be certain to let everyone know. I don't leave threads like that.

The key points about how this live culture was treated was:

1) I used the 4% malt/dextrose mix which many claim is among the best mixes
2) The solution was incubated at 82-84F the entire time (I figure this is at least as important as any other factor).
3) The solution was constantly stirred
4) I started 1ml of stored live culture, not spores.

It could have been dumb luck or something. I have no idea. Like I said, I will be sure to write back if this stuff is green mold.

PS - it's been injected into a sterile mix of 50% horse manure and 50% WBS, supplemented with about a small amount of stevia leaf, bee pollen, and tiny bit of black mustard seed. The mixture's moisture was adjusted with vermiculite.


Edited by Blue Helix (06/27/05 11:35 AM)


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OfflineBlue Helix
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Re: Constant-stir LC showing expotential growth in first 3.5 days. (pics) [Re: debianlinux]
    #4342415 - 06/27/05 11:37 AM (11 years, 10 months ago)

I had been reading about week to two week development times, but maybe this is normal. What's the norm for about half the jar being a mycelium cloud?


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Invisibleagar
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Re: Constant-stir LC showing expotential growth in first 3.5 days. (pics) [Re: Blue Helix]
    #4342463 - 06/27/05 12:05 PM (11 years, 10 months ago)


Tough thing, is there is no normal.

All strains differ slightly (miniscule amount).

As does the nutriant source & percentage.

As does the jar size & amount of solution.



This is what I get after 4 to 6 days.



This is 6 to 7 days & time to place it in the fridge.


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OfflineBlue Helix
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Re: Constant-stir LC showing expotential growth in first 3.5 days. (pics) [Re: agar]
    #4342485 - 06/27/05 12:18 PM (11 years, 10 months ago)

This morning was 4 days for me and mine was between your bottom left and the top ones. I never let mine settle as far as the bottom right. In the bottom left case, your mycelia aggregates look larger than the ones in my liquid culture. I wonder if that is what the constant stirring did, just reduce the size of the aggregates.

Really the only way to tell if the constant stir thing helps or not is to run two jars with identical solutions and identical inoculations side by side, keeping both at the exact same temperature but stirring one constantly and the other daily. Otherwise, there are just too many variables.

I just wanted to show what I got from the constant stirring. I was amazed at the speed of growth. I certainly did not expect to use it on day four like I did.

By the way, I got this 140 ml syringe for $10. When I bought it I thought it was a rip off because it was so expensive, but I really wanted one larger than the cheap 60 ml ones. But that 140 ml syringe REALLY kicked ass for using the LC! I could inject each spawn bag with a hefty 140 ml. And the super large syringe allowed me to suck up 140 ml with a huge needle, change to a smaller needle, settle some of the mycelium in the syringe for a few minutes, and inject the concentrated cloud into the vaccutainers with the small needle. I just worked so perfectly! Those huge syringes are nice.


Edited by Blue Helix (06/27/05 12:20 PM)


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Invisibleagar
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Re: Constant-stir LC showing expotential growth in first 3.5 days. (pics) [Re: Blue Helix]
    #4342686 - 06/27/05 01:20 PM (11 years, 10 months ago)


:wink: :tongue:


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Invisiblemycofile
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Re: Constant-stir LC showing expotential growth in first 3.5 days. (pics) [Re: agar]
    #4344825 - 06/27/05 10:38 PM (11 years, 10 months ago)

Blue Helix, I don't see your syringe, is it the one agar posted? Where could that be found? I've seen similar at vet supply sites, but never used one. I assume it's autoclavable?


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Re: Constant-stir LC showing expotential growth in first 3.5 days. (pics) [Re: mycofile]
    #4345254 - 06/28/05 12:53 AM (11 years, 10 months ago)

Quote:

mycofile said:
Blue Helix, I don't see your syringe, is it the one agar posted? Where could that be found? I've seen similar at vet supply sites, but never used one. I assume it's autoclavable?




That one is a Socorex (made in Switzerland) & is fully autoclaveable.

http://www.socorex.ch/scripts/Eprogresultfin.asp?cat=163&langue=E

I snagged it new in factory box off eBay for $26 (about 1/6th of retail).

It is a smooth working & easy to use SOB. Uma @ MushMush uses the same one.


Edited by agar (06/28/05 03:43 AM)


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OfflineBlue Helix
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Re: Constant-stir LC showing expotential growth in first 3.5 days. (pics) [Re: agar]
    #4345440 - 06/28/05 02:02 AM (11 years, 10 months ago)

Mine is nothing fancy like agar's. It's a simple 140cc Monoject like this:

http://lambriarvet.com/sbsite.php?&item=01101MON-140LS

Of course they are autoclavable. I bought a 2" 16 guage needle to go with it. I have another 20 guage needle for the more fine work of injecting into the silicon stopper once the lc settles in the syringe since the 16 guage really beats up a septum real bad.


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Re: Constant-stir LC showing expotential growth in first 3.5 days. (pics) [Re: Blue Helix]
    #4345533 - 06/28/05 02:30 AM (11 years, 10 months ago)

where do you get the big, long needles?


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OfflineBlue Helix
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Re: Constant-stir LC showing expotential growth in first 3.5 days. (pics) [Re: Blue Helix]
    #4345616 - 06/28/05 02:51 AM (11 years, 10 months ago)

I looked at that big needle again. I guess it's bigger than I thought. It's actually a 2" x 12 guage needle. I have a local lab supply that sells 2" 12 guage blunts for about $1.50. They also sell 16-guage 8" blunts for $4. I can't do much with 16-guage except use them to inject the septum.

12 guage is just about perfect for a big 140CC syringe and using live cultures. That way you can inject the whole 140ccs into your spawn bag in one sitting and hot glue it shut immediately. Before they go in the PC, I always use duct tape covered with masking tape on my spawn bags. The duct tape provides strength but melts so I protect over it with the less strong but non-melting masking tape (so it doesn't stick to the walls of the pressure cooker). Those two make a strong injection base and good surface for the hot glue to seal the hole:



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Re: Constant-stir LC showing expotential growth in first 3.5 days. (pics) [Re: Blue Helix]
    #4348965 - 06/29/05 02:02 AM (11 years, 10 months ago)

Hmmm. That's pretty quick. I don't own a stir plate, so it's hard for me to say. I've been using vanilla in mine and they're ready to go in 6-8 days. If! Maybe 4-5 with a plate. But 6-8 with no stirring capabilities. There's a big difference in a properly stirred LC with a plate and hand swirled one for sure. I can buy it though.


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OfflineBlue Helix
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Re: Constant-stir LC showing expotential growth in first 3.5 days. (pics) [Re: Hotnuts]
    #4349079 - 06/29/05 02:33 AM (11 years, 10 months ago)

It's been 24 hours and the spawn bags look like they are perfectly inoculated. I see tiny white spots all over evenly spread and they look exactly like cubensis mycelium, not mold as I feared might happen. At this rate, I predict the bags will complete and be white as paper in 4 more days. If my timing is right, that would mean I went from 1 ml mycelium solution in a syring (1 gram) to 25 pounds of colonized spawn (11350 grams) in 9 days. That is an increase of mycelium-infected substrate of four orders of magnitude in 9 days -- simply amazing! If I could grow that fast, I'd probably be the size of small country in a couple weeks! lol


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Re: Constant-stir LC showing expotential growth in first 3.5 days. (pics) [Re: Blue Helix]
    #4349418 - 06/29/05 04:10 AM (11 years, 10 months ago)

nice, update us with some pics on the bags please.
peace ohm :mushroom2:


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OfflineBlue Helix
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Re: Constant-stir LC showing expotential growth in first 3.5 days. (pics) [Re: ohmatic]
    #4349942 - 06/29/05 11:18 AM (11 years, 10 months ago)

It has been 48 hours since the LC inoculation:



At this rate, I believe these will be done in no more than 72 hours bringing the total to 5 days for colonization.


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Mushrooms, Mycology and Psychedelics >> Advanced Mycology

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