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Pastywhyte
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Registered: 09/15/12
Posts: 37,972
Loc: Canada
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Re: Rye Soak Water Liquid Culture! [Re: Violet]
#21041891 - 12/29/14 10:53 PM (10 years, 20 days ago) |
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I did brown rice water LC a few months ago. Worked good but there was a lot of sediment. Myc tore through it pretty nice tho.
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Goldberg
Stranger


Registered: 08/31/14
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Re: Rye Soak Water Liquid Culture! [Re: Pastywhyte]
#21042977 - 12/30/14 08:37 AM (10 years, 20 days ago) |
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This is fučking awesome.. Sometimes you do so much research that you forget shit- really important shit like this You even tried to tell me about GWLC last night pasty and I completely overlooked it as I get tiresome of acronyms and will sometimes jump right over them while I'm reading something When I first got into agar I was having so much difficulty using LME agar that I started using GW instead of LME for my agar mix- and I had a breakthrough Since then I switched back to lme for agar plates which is why I didn't think to use GW for the LC I've been making... yay for grainwater
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Violet



Registered: 12/06/11
Posts: 4,205
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Re: Rye Soak Water Liquid Culture! [Re: Goldberg]
#21042993 - 12/30/14 08:42 AM (10 years, 20 days ago) |
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Quote:
Goldberg said: This is fučking awesome
yay for grainwater
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hamloaf
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Registered: 12/23/09
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Re: Rye Soak Water Liquid Culture! [Re: Violet]
#21043304 - 12/30/14 10:12 AM (10 years, 20 days ago) |
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That's cool that shroominmyroom is conducting their own research with their grain boil water.
I wouldn't have strained off the sediment. 9 hours to strain the medium, to me, defeats the purpose of utilizing this culturing medium as a means to save time, money, and effort spent on gathering and preparing the ingredients to make the LC. Sediment in liquid media is a good thing for the liquid myce. Sediment gives the dikaryotic mycelium strands something to grab onto, feed from, and leap-off of.
The only reason I would choose to dilute was to get faster colonization. Other than that, I prefer media with high nutritional content. So does the mycelium. Plan to add nutritional yeast to another non-diluted rye/grain boil water liquid culture test batch, as well as try the diluted formula with, and with out the addition of nutritional yeast. I have a clone culture being worked on down to an isolate that's on it's third transfer.

Once I hit a monoculture, the plan is to conduct the next round of liquid culture experiments with said isolate, and see what's really going on. This medium is SHREDDING the grains. Only 6 days from inoculation and these jars are at over 25% colonization, and could have been shook yesterday.
Shook em' up this morning.

It's being projected that these grain cultures will have their medium colonized with in 2-5 days from the shake.
Soaking some high quality Red Milo right now. Makes sense that I should whip up a few LC's using the Milo boil water, so I am posting about this in hopes that I don't forget to do exactly that in the midst of the huge workload of today.
Two things I am not sure of: 1. How to dilute the medium. 2. Whether or not doing a Milo Boil Water Liquid Culture warrants it's own thread.
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Pastywhyte
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Registered: 09/15/12
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Loc: Canada
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Re: Rye Soak Water Liquid Culture! [Re: hamloaf]
#21043317 - 12/30/14 10:16 AM (10 years, 20 days ago) |
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Keep it in this thread ham. Looking boss tho, I am glad its working as well for you as it did for me 
When I dilute I go no more than one part gw to four parts water. I find that anything in between also works but that is the most I stretch it.
Edited by Pastywhyte (12/30/14 10:21 AM)
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blackdust


Registered: 02/28/09
Posts: 8,327
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Re: Rye Soak Water Liquid Culture! [Re: blackout]
#21043841 - 12/30/14 12:35 PM (10 years, 20 days ago) |
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Quote:
blackout said: They might be doing better as they are used to the substrate
This is from Stamets GGMM talking about agar
Quote:
The pH of the above media formulations, after sterilizing, generally falls between 5.5-6. 8. This media can be further fortified with the addition of 3-5 grams of the end-substrate (in most cases hardwood sawdust) upon which mushrooms will be produced. If samples of soil or dung are desired, they first must be pre-boiled for 1-2 hours before adding to any of the above formulas. One potential advantage of the addition of these end substrate components is a significant reduction in the "lag period". The lag period is seen when mushroom mycelium encounters unfamiliar components. (See Leatham & Griffin, 1984; Raaska 1990). This simple step can greatly accelerate the mushroom life cycle, decreasing the duration of colonization prior to fruiting.
I have been thinking about this statement by him and other like this everyday for the past few weeks now.
--- hamloaf, thank you for the detailed reports and pictures. You are sharing some amazing value to the community. You need to be promoted to a TC. You have been giving good advice for over 5 years now. I know I have learned from you 5 years ago and still to this day. 5 shrooms to you!
Edited by blackdust (12/30/14 12:37 PM)
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hamloaf
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Registered: 12/23/09
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Re: Rye Soak Water Liquid Culture! [Re: blackdust]
#21047564 - 12/31/14 07:03 AM (10 years, 19 days ago) |
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Round 2, FIGHT! Made some liquid media out of some high quality Red Milo water from water at the end of the boil cycle yesterday afternoon. One media was was diluted down to 4%, and the other was not diluted. 10-15 kernels were added to each media. The kernels burst under the intense heat of the sterilizer creating more sediment in the media which is preferred by this cultivator. No nutritional yeast was added.

The mediums were then sterilized for 45 minutes at 15-17 psi.

The plan is to inoculate these tomorrow with some of that clone transfer (T2, or T3) culture when transfers from those plates to more agar are made. Plan on inoculating several several grain masters with the culture not selected for further sectoring as well.
The mycelium is expected to colonize the diluted medium slightly faster than the undiluted one, because when a substrate or medium has lower nutrition in it, the myce is forced to go in search of food resulting in faster colonization. The more nutritious the substrate is causes the food source for mycelium to be abundant enough so that the myce doesn't have to feverishly hunt for more food resulting in a slightly slower colonization rate.
The phenomenon of the myce colonizing more nutritious substrates, or culturing media slower than substrates or culturing media due to excess nutrition is something that is usually/more often than not made up for at fruiting time.
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Pastywhyte
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Registered: 09/15/12
Posts: 37,972
Loc: Canada
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Re: Rye Soak Water Liquid Culture! [Re: hamloaf]
#21047596 - 12/31/14 07:29 AM (10 years, 19 days ago) |
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Sweet. Should be a great thing to watch. I bet the 4% still does well, better than 2% sugary LC would
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shroominmyroom
Grasshopper


Registered: 10/30/14
Posts: 3,639
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Re: Rye Soak Water Liquid Culture! [Re: Pastywhyte]
#21048401 - 12/31/14 11:56 AM (10 years, 19 days ago) |
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Edited by shroominmyroom (07/31/17 11:26 AM)
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taGyo
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Registered: 10/16/14
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So in order to create a GWLC can I just take some of my oats boiling water and put it in a jar and sterilize then inoculate?
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hamloaf
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Registered: 12/23/09
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Re: Rye Soak Water Liquid Culture! [Re: taGyo]
#21053242 - 01/01/15 05:24 PM (10 years, 17 days ago) |
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Yes. As stated above, you could dilute down to 4%, or any where in between, or you could just use straight, non diluted boil water.
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taGyo
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Re: Rye Soak Water Liquid Culture! [Re: hamloaf]
#21053390 - 01/01/15 06:03 PM (10 years, 17 days ago) |
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Thanks Ham,
Will post up some pics.
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hamloaf
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Re: Rye Soak Water Liquid Culture! [Re: taGyo]
#21057524 - 01/02/15 05:17 PM (10 years, 16 days ago) |
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I am disgraced to report that 2 of the grain cultures were lost to what appears to be bacillus. You can tell by the greasy appearance of the grains without any mycelium colonization.
My guess as to where the contamination came from was probably in my sterile technique for that day. I was getting too pictious and most likely cross-contaminated somewheres. Solution: Stop using the camera so much during lab work.
The other 4 grain mediums are doing well, however.

Give them another 3-4 days tops before the jars become fully colonized.
Got the Milo Boil Liquid Mediums inoculated, as well, with some clone culture on it's 3 transfer.

More to come as developments are made. Thanks for checking this out.
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azur
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Registered: 04/21/12
Posts: 28,103
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Re: Rye Soak Water Liquid Culture! [Re: hamloaf]
#21057832 - 01/02/15 06:30 PM (10 years, 16 days ago) |
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Good stuff ham. Funny how things have changed in people's views
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13shrooms
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Registered: 01/01/09
Posts: 26,719
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Re: Rye Soak Water Liquid Culture! [Re: hamloaf]
#21059277 - 01/03/15 02:01 AM (10 years, 16 days ago) |
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Quote:
hamloaf said:
There is a correlation here going on between cutting out the sugars in LC, and only using light malt extract, and better performance of the culture that I can't explain.
Bulk subs made from LC's with sugar, ime, haven't done the best at fruiting either. 
Knowing how much liquid mycelium to inoculate any substrate with that equals, or would be similar to inoculating a substrate with a high grain ratios would help too I think.
thats what Ive been saying for a bit now, its anaphylactic shock and why TL and I wrote this LC tek and use high spawn ratios 
http://www.shroomery.org/forums/showflat.php/Number/15234205
http://www.shroomery.org/forums/showflat.php/Number/14722933
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hamloaf
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Registered: 12/23/09
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Re: Rye Soak Water Liquid Culture! [Re: 13shrooms]
#21059405 - 01/03/15 03:50 AM (10 years, 16 days ago) |
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Yea, I know, but I want to know what the equivalent of inoculating grain media with liquid mycelium that equals inoculating grain media with high spawn ratios. You dig?
This is the major reason that I am getting sick of doing LC's by hand again. You can just never be sure of the concentration of liquid mycelium at any given time. That's why the only way I see to making LC's worth carrying on developing with (for this cultivator's individual needs and growing situation) is to get a Eberbach, a stir plate, and few stir bars.
PS ~ Already have an Eberbach Waring blending unit, and 500ml glass container with lid on it's way and should be here any day now. Been looking around for/at stir plates. There are a lot of options out there. Some say that they are not for continuous use. Others say they have a "hot plate" option, which is basically a heating element under the stir plate that heats up. I am just not sure which type of stir plate(s) best suit the needs of this hobby, so I have refrained from getting one.
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taGyo
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Registered: 10/16/14
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Re: Rye Soak Water Liquid Culture! [Re: hamloaf]
#21059874 - 01/03/15 09:12 AM (10 years, 16 days ago) |
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First two GWLC,
Got 4 more sitting. Inoculated with B+ and GT spores I had laying around. Pretty dark. Watered down two of them, sterilized for an hour, gave them a good shake and then inoc'ed.
-------------------- Gyo's Better Grows
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Dominus fortunae meae sum
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shroominmyroom
Grasshopper


Registered: 10/30/14
Posts: 3,639
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Re: Rye Soak Water Liquid Culture! [Re: taGyo]
#21059943 - 01/03/15 09:37 AM (10 years, 16 days ago) |
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Edited by shroominmyroom (07/31/17 11:30 AM)
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Pastywhyte
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Registered: 09/15/12
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Re: Rye Soak Water Liquid Culture! [Re: taGyo]
#21059949 - 01/03/15 09:39 AM (10 years, 16 days ago) |
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I have disliked sugar based LC for some time now. The SITR LC that I decided to try has reinforced my stance yet again. The myc is slow to recover and I am going to go back to storing my grain soak water again for my next LC. I am also giving malt a go again, hopefully that won't let me down short term, never tried malt before for LC but my last jars worth of grain water cracked in the PC so I had no choice.
Regardless in terms of recovery time and the myceliums performance after, I like gwlc a lot. Its a little tricky to deal with the sediment etc but once you get those things figured out I love the performance.
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taGyo
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Registered: 10/16/14
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Re: Rye Soak Water Liquid Culture! [Re: Pastywhyte]
#21059982 - 01/03/15 09:49 AM (10 years, 16 days ago) |
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Quote:
shroominmyroom said: I'd like to suggest agar wedge for inoculation. It's dangerous to use spores from syringe.
I'm guessing your LC was made from oats. It does look rich. What's the ratio of GW to water?
I like to fill the jars up half way. It's easier to swirl LC around without touching the filter.
Do you happen to know if syringe filters are water proof?
Water proof and autoclavable. I don't even put foil on top of these jars, just throw 'em in the PC with the ring loosened a smidge. The one time I didn't loosen the ring I pulled out my LCs and heard "Tssssssssss" like a ticking time bomb.
Let that bitch cool in the next room.
One I put about 1/2 water, 1/2 GW, another I put 1/4th water and rest GW. No matter how much water I put though they all came out extremely murky.
Oats would be correct.
I'm gonna throw some more on today and try to get it so it's not that murky and inoc with some APE.

Quote:
Pastywhyte said: I have disliked sugar based LC for some time now. The SITR LC that I decided to try has reinforced my stance yet again. The myc is slow to recover and I am going to go back to storing my grain soak water again for my next LC. I am also giving malt a go again, hopefully that won't let me down short term, never tried malt before for LC but my last jars worth of grain water cracked in the PC so I had no choice.
Regardless in terms of recovery time and the myceliums performance after, I like gwlc a lot. Its a little tricky to deal with the sediment etc but once you get those things figured out I love the performance.
I remember you telling me about the SITR LC and I was wondering how that was going versus GWLC.
The thing about GWLC is I don't understand why more people DON'T do it. It's easy LC water after I've prepped my grains and you can just throw them in the PC with your grain jars and be completely fine. I'm gonna take a colonized grains from two jars that have bacteria in 'em and drop it in these LCs.
Thanks for the heads up Ham and Pasty,
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