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One MS inoculated LC split into two


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#1 RutgerHauer

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Posted 11 February 2019 - 02:03 PM

So, recently, when the thought entered my mind I had been reviving my hobby with a dead culture in a liquid medium stored away in inconsistent fridge temperatures - for way too long - I though I might as well make a big batch now I know how to do it myself. That way I don't have to buy a lousy 25 ml vial of sterilized honey water for 25 euros, as I did when I started out.

 

Some time around Christmas I got a jar of 440 ml and made 400 ml of liquid culture medium by adding 16 grams of maple sirup like stuff to tapwater, about 4% in weight (a little less) and sterilized it properly. But - I was just getting back into informing myself online and just a few days after I inoculated the jar - and came upon the thing called a Self Healing Injection Port - Dang, I should have known this before!

 

So, got on it, I figured out a local source to get some RTV silicon and ordered a bunch of new jars and fixed them up crudely but functionally after some trial and error. So - what to do about the culture that is 1 - way too much for one jar and 2 - has no SHIJ? Improvised an SAB, got extra cleanly with stuff after I had prepared two jars with 50 ml of the same honey water mixture as I did before and simply divided the 400 ml culture into the two 500 ml jars filled with 50 ml of LC. Now I have two jars of 'selected' mycelium and spores:

 

I did it in such a way, that I very gently swirled the jar so only the light parts of germinated mycelium and probably spores, floated up to the top. That went into jar A on the 11th of January, and what was left in jar 0 on the bottom went into jar B on the same day (like a few seconds later, lol).

 

I am now working with jar A - since that was the mycelium that had germinated the quickest. I used it a day later with minimum growth visible in the liquid, but it started colonizing my grains in two to three days. Very nice. I am curious about jar B though. In the mean time it has developed a bit more but seems to be less happy and healthy than their nephews in jar A. They tend to form more like 'knot'-like structures and beef up in that way, rather than the more spread out growth of the mycelium in jar A. You might also be able to see the colour difference between the LC's - jar B is a bit darker/yellower as it probably has not used up as many sugars in the water.

 

Will have to see in the future what and or there are in fact any differences at all between the two. Will update on that later for sure!

RH

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Edited by RutgerHauer, 11 February 2019 - 08:22 PM.

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#2 RutgerHauer

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Posted 11 February 2019 - 02:05 PM

I try to look at this picture cross eyed to make in 3D but it doesn't work.

 

 

Something else - I read on that other Shroom forum someone saying add 2% agar agar to your LC for extra succes - I'm not thinking about doing this because just honey or sirup works fine - and won't this just make it a jar full of agar? lol. Is it like a 'joke' to say stop doing LC and go Agar?


Edited by RutgerHauer, 11 February 2019 - 02:18 PM.

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#3 onediadem

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Posted 11 February 2019 - 09:19 PM

I freaking love the speed lc's provide! I hope they are clean!


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#4 Billcoz

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Posted 12 February 2019 - 01:12 AM

Nice work! I made some clear karo syrup LC jars the other day, gonna bang em up with tiny tissue/myc samples from agar dishes I have, one colonized with B+, and one burma, each have been isolated, not quite down to a single-sector strain, but close. Doing the inoculations in the morning.


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#5 RutgerHauer

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Posted 12 February 2019 - 10:59 AM

I freaking love the speed lc's provide! I hope they are clean!

 

That's the big question of course.. All colonized jars look healthy - but I guess nothing is sure until I have a flush going.

 

Nice work! I made some clear karo syrup LC jars the other day, gonna bang em up with tiny tissue/myc samples from agar dishes I have, one colonized with B+, and one burma, each have been isolated, not quite down to a single-sector strain, but close. Doing the inoculations in the morning.

That's the way to go, good luck with that!



#6 RutgerHauer

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Posted 18 February 2019 - 05:04 PM

Has been a few but I have had not much to share; still waiting on my first tray to fruit and waiting on eight 440ml jars of popcorn to colonize. In the mean time I am getting the last bits and pieces of my setup together so I'll be set and ready to go anytime in the future.

Something 'interesting' happened with the latest run from LC2A; on one jar I probably shot a blank in there. Growth would hardly get going in jar A and decided to shake it even though it was far behind from the rest. Recovery was bad and shook it again after a week in a final attempt to help it spread, but I think it has stalled now. Still not bad, seven out of eight jars getting there!

Picture below shows this jar a few days after the last shake (basically uncolonized corn) - and jar H, which I did not shake at all because it went so quick it would make it on its own. All other jars after a shake and a week are at a similar stage of colonization now as jar H at about 95%.

RH

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Edited by RutgerHauer, 18 February 2019 - 09:50 PM.

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#7 RutgerHauer

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Posted 21 February 2019 - 11:25 AM

I have pins today! Yesterday broke up 7 jars of corn and put into a second tray. No photo's, just excitement!
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#8 RutgerHauer

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Posted 25 February 2019 - 04:11 PM

I have this going on now. Not sure what is happening, mutants, too little of a substrate, low humidity.. I guess I'll just wait and see.

My confusion is that it are pins with open caps. Will those continue growing or stay small? Not sure. I probably made a mistake in choice of the shallow layer of grain substrate with a thicker casing of vermiculite. Could be I didn't hydrate the casing layer enough, coming to think of it. This field capacity thing..

What do you guys think? I think it's cute.

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#9 Billcoz

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Posted 26 February 2019 - 01:10 AM

 

I get some like that sometimes, I think from too much fae and there could be genetics that do that. They are cute, and IMO they can be potent. They should continue to grow, the caps seem to not have veils but they will open and drop spores.


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#10 RutgerHauer

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Posted 26 February 2019 - 06:46 AM

I get some like that sometimes, I think from too much fae and there could be genetics that do that. They are cute, and IMO they can be potent. They should continue to grow, the caps seem to not have veils but they will open and drop spores.


Thanks for the reply. I had concluded the same. Had been reading the importance of FAE to help the mushrooms pinning, but I might have overdone it - good lesson for the next time. Since yesterday I dialed it back a lot and that upped the humidity.

Edited by RutgerHauer, 26 February 2019 - 10:13 AM.


#11 RutgerHauer

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Posted 26 February 2019 - 10:35 AM

A day later.

This is a cooperation of two separate inoculation dates, about a week apart, might explain the uneven pin set!

Next one will go into FC today, fromthe same incolulation date and with a substantial amount more grain as a substrate.

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Edited by RutgerHauer, 26 February 2019 - 10:47 AM.

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#12 RutgerHauer

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Posted 27 February 2019 - 08:11 AM

I think I will harvest some today. I think they stay small on this run, which is fine!

Edit: I took just under 35 grams wet off of the tray. Pretty stoked I'm finally seeing results, took long enough!

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Edited by RutgerHauer, 27 February 2019 - 09:09 AM.


#13 RutgerHauer

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Posted 28 February 2019 - 07:52 AM

Today another 25 grams. I like it.
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#14 Deleena24

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Posted 28 February 2019 - 04:47 PM

What's the strain Rutger? May have something to do with the funny fruits.
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#15 RutgerHauer

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Posted 28 February 2019 - 05:05 PM

What's the strain Rutger? May have something to do with the funny fruits.


Excuse me, I thought I had mentioned, but I told everything except this, there's just GT on my liquid cultures which stands for Golden Teacher. They grow a bit differently from when I grew some in a growkit a while ago. That's just different fenotypes since I grew from MS I guess. Also, circumstances make it logical for the mycelium to produce smaller fruits I think, like when a mushroom grows on a petridish.
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#16 RutgerHauer

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Posted 01 March 2019 - 10:39 AM

On the third day I conclude the 'first flush' is finished; another 28 grams wet makes a total of 88 grams of fresh mushrooms. Somehow I was expecting less for a first grow.

I see the substrate is starting shrink on the sides of my tray. Is this a good time to add some water or is misting sufficient?

#17 Billcoz

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Posted 01 March 2019 - 12:43 PM

If it feels lighter than you would expect from the thickness, and what it felt like when you first madeit, then you can soak it for a few hours, I wouldn't go over 12, i do 8 generally, or sometimes a quick 2 hours or so. I like to remove any pins that are left if any, they tend to abort. The sub will float so you can weigh it down with something, some people just float it Be careful though, it will crumble or crack easily. The best way to do it if your tray isn't deep enough to submerge it is to set the whole tray into a deeper tray or bin, then fill it and when it overflows fill the bigger one, that way you do not have to remove it from the tray, making it less likely to break or damage it. Oh, and a good way to drain it is to lay something wider and longer than the tray that is flat like a board over top to cover it, hold that down on the sub to hold the sub it the tray as you tyrn it upside down and drain, that way the sub don't fall out of the tray when you are draining it.


Edited by Billcoz, 01 March 2019 - 12:47 PM.

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#18 RutgerHauer

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Posted 01 March 2019 - 12:59 PM

I'm not sure a complete dunk is the right choice here, or at least now. I think I'll check back tomorrow and see how's things are going. It's tricky since pins are coming up very randomly and pretty much continuously.. Can't harvest everything at once that way.

How about side watering? I know for a fact he mushrooms have taken up about 80 ml of water, but that of course would not take in count any other use of water by the mycelium and evaporation.. So I would not be sure how much water to add.

Edited by RutgerHauer, 01 March 2019 - 01:00 PM.


#19 Billcoz

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Posted 01 March 2019 - 01:08 PM

I'm not sure a complete dunk is the right choice here, or at least now. I think I'll check back tomorrow and see how's things are going. It's tricky since pins are coming up very randomly and pretty much continuously.. Can't harvest everything at once that way.

How about side watering? I know for a fact he mushrooms have taken up about 80 ml of water, but that of course would not take in count any other use of water by the mycelium and evaporation.. So I would not be sure how much water to add.

Yeah that might be a good call, it is not always necessary, but at some point, maybe after another flush or two, it will feel light enough to notice, so lift it everyday to make sure. My current tubs just needed a dunk after the first flushes, they were big flushes so I wasn't surprised, they were very light for the depth of sub.


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#20 RutgerHauer

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Posted 01 March 2019 - 01:39 PM

I also made up my mind for the next run. I was thinking about making a switch in grain to make spawn. I am using corn now with success, but want to try millet. I quickly realized after buying a kilogram of the stuff it is quite expensive if you want to make a lot of jars since it is such a dense/compact grain.

I am still using it to colonize, but with less jars, to do a grain to grain transfer to prepared corn jars. I figure the millet will fill up the spaces in between the corn kernels pretty well, and would make a nice mixture of the two - and for the bigger part of my grains I will still be able to go on cheap popcorn.

Edited by RutgerHauer, 01 March 2019 - 01:44 PM.





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