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Offlineinked4life
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Multi spore syringe and questions on monokaryotic mycelium
    #21955231 - 07/17/15 08:59 AM (8 years, 6 months ago)

Recently I inoculated rye grain berries with a ms syringe. All the growth has been fuzzy white mycelium and no rhizomorphic  growth. I am wonder if it's possible to have a monokaryotic  mycelium  from a ms  syringe. I would think with having multi spores germinating and being shook up that it would have found mating mycelium that would produce fruits. This has not been accomplished yet. I have also isolated on pda different grains from different parts of the spawn. Taken 2 separate grains on 1 dish and tried to create rhizomorphic  growth. To early to tell on the pda  but all of the dishes 8 in all look to have the same fuzzy consistent  mycelium. Could it be the vendor or the spores I started off with.


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Invisibleblindingleaf
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Re: Multi spore syringe and questions on monokaryotic mycelium [Re: inked4life]
    #21955281 - 07/17/15 09:17 AM (8 years, 6 months ago)

monokaryotic=single spore germination

MULTI spore syringe = more than 1 spore.

technically, i guess all the spores could have died but one…but chances are slim to none.  monokaryotic growth is slow and hard to see with naked eye.


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Invisibleinsanemike

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Re: Multi spore syringe and questions on monokaryotic mycelium [Re: blindingleaf]
    #21955317 - 07/17/15 09:30 AM (8 years, 6 months ago)

Op, choose the fastest growing growth from each of those plates and transfer them to as many plates as you can. Keep repeating this process until you have one single strain on each plate. Test them all on a small scale and choose the most prolific strain to slant for long term storage so that you have a base from which to start each grow.

tip: cut back on the nutrional content in your agar, the mycelium won't grow as thick and fluffy.


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Offlineinked4life
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Re: Multi spore syringe and questions on monokaryotic mycelium [Re: insanemike]
    #21955361 - 07/17/15 09:45 AM (8 years, 6 months ago)

how do i cut down on the nutrients. i ordered my pda off of ebay thru some china supplier. just comes in a bag. had to look up how much destilled water to use per gram of pda. no directions were included. i used 500 ml destilled water to 19.5 grams of pda. set up nice and took the transfers great. just keep repeating ay?


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Invisibleinsanemike

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Re: Multi spore syringe and questions on monokaryotic mycelium [Re: inked4life]
    #21955750 - 07/17/15 11:37 AM (8 years, 6 months ago)

Yes. Keep transferring but only take the fastest, most outer edge of growth each time. You can get away with about 2.5g per 100 ml of water. I don't use agar that already has added nutrients and you probably won't anymore either once you know what you are doing.


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InvisiblebodhisattaMDiscordReddit
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Re: Multi spore syringe and questions on monokaryotic mycelium [Re: insanemike]
    #21955801 - 07/17/15 11:52 AM (8 years, 6 months ago)

To get monokaryotic growth you would usually do something like dilute your spore syringe 1:100 and then squirt a single drop onto an inoculation loop and make a streak plate.


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Offlineinked4life
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Re: Multi spore syringe and questions on monokaryotic mycelium [Re: bodhisatta]
    #21955876 - 07/17/15 12:12 PM (8 years, 6 months ago)

i just never seen mutliple germination spots create the same type of fuzzy mycelium that doesnt have any rhizo growth. Has me questioning everything from each tek i used. My colonization time was very fast, no contamination but no fruiting at all. That is why i thought maybe monokaryotic because of no pins and no rhizo growth. I got the supplier to send me something off their inventory so hoping to get some more spores soon. Think i might go back to brf cakes since grains havent been successful, ive had great success with cakes in the past. and im using a sgfc with polyfill instead of 1/2 holes drilled into a tub and using a cool misting unit.


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InvisiblebodhisattaMDiscordReddit
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Re: Multi spore syringe and questions on monokaryotic mycelium [Re: inked4life]
    #21956038 - 07/17/15 12:58 PM (8 years, 6 months ago)

Noobs are for whatever reason astoundingly over concerned with rhizo growth. Tomentose cultures can be just as good or better. Worry about clean growth not how it closes to grow.

Sounds like your I know better than years of trial and error fruiting chamber is the problem to be honest


Edited by Trusted cuItivator (07/17/15 12:58 PM)


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Invisibleinsanemike

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Re: Multi spore syringe and questions on monokaryotic mycelium [Re: bodhisatta]
    #21956130 - 07/17/15 01:18 PM (8 years, 6 months ago)

I have to agree with bod that rhizo is more of an aesthetic type thing. If you are looking for a mono culture that is a definite fruit producer, you should clone a fruit from one of your pf cakes and then isolate from there. But like bod said, genetics may not be your culprit, your fruiting chamber/conditions may be.


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InvisibleMad Season
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Re: Multi spore syringe and questions on monokaryotic mycelium [Re: insanemike]
    #21956136 - 07/17/15 01:19 PM (8 years, 6 months ago)

The moment I hear a cool mist setup with polyfil, I can't help but think it's a chamber problem. Gizmos will never be able to replace your eyes.


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Offlineinked4life
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Re: Multi spore syringe and questions on monokaryotic mycelium [Re: Mad Season]
    #21959551 - 07/18/15 07:54 AM (8 years, 6 months ago)

no i ditched the cool mist with the last fc , i havent used it. just fanning couple times a day. i transferred to pda what healthy myceluim i had and planted my casing that didnt pan out outside. my issue is misting. i dont know how much is to much or if im just doing too little.


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Offlineinked4life
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Re: Multi spore syringe and questions on monokaryotic mycelium [Re: inked4life]
    #21959555 - 07/18/15 07:56 AM (8 years, 6 months ago)

i often see casing that are outside of a fruiting chamber . is that ok with casing.


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Invisibleinsanemike

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Re: Multi spore syringe and questions on monokaryotic mycelium [Re: inked4life]
    #21959664 - 07/18/15 08:40 AM (8 years, 6 months ago)

When you use the word casing, are you referring to cased grains?


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Offlineinked4life
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Re: Multi spore syringe and questions on monokaryotic mycelium [Re: insanemike]
    #21959893 - 07/18/15 10:13 AM (8 years, 6 months ago)

yes, grains .


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Offlinespacechildo
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Re: Multi spore syringe and questions on monokaryotic mycelium [Re: inked4life]
    #21959905 - 07/18/15 10:19 AM (8 years, 6 months ago)

the casing layer goes on top of grains or your bulk sub either right into the fc
or in a tray which you then put in a fc.


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Invisibleinsanemike

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Re: Multi spore syringe and questions on monokaryotic mycelium [Re: spacechildo]
    #21960035 - 07/18/15 11:04 AM (8 years, 6 months ago)

Grains can be difficult to fruit because it's easy to over and under mist. The casing can help provide a nice micro climate but will easily dry out if neglected for more than a day. Bulk substrates help keep humidity high and therefor make them more of a set and forget. Never heavily mist cased grains, even if you let the casing dry out for too long.


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Offlineinked4life
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Re: Multi spore syringe and questions on monokaryotic mycelium [Re: insanemike]
    #21960523 - 07/18/15 01:01 PM (8 years, 6 months ago)

i used jiffy mix 50/50 peat moss and verm and a lil lime or something. pc'd it for 60 mins at 15 psi. no contams but all the casing dried out turn blue as shit so i buried them outside and isolated one. took samples and transferred to pda. my spore supplier is sending me more spores also. i am going to go back to cakes. anyone heard of using rye grains and verm for cakes. or should i stick to the pf tek brf for cakes, i like how grains colonize so i was wondering if you can use pint wide mouth jars and mix verm with prepared berries and treat them as cakes. would they fall apart if you tried dunking them. brown rice flour is just harder to get here.


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Invisibleinsanemike

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Re: Multi spore syringe and questions on monokaryotic mycelium [Re: inked4life]
    #21960666 - 07/18/15 01:36 PM (8 years, 6 months ago)

Stick to the pf tek. Do some reading up on bulk substrates and proper methods before moving on to bulk. The mycelium can have a blue tint when they are stessed but that doesn't necessarily mean that they are contaminated. The casing can and will turn green with trich if you sterilize it the way that you did. Sterlization and pasteurization are 2 different methods. Sterilization wipes out almost all microbes and gives mold spores the window of opportunity to germinate and grow where as proper pasteurization kills mold and bacteria leaving behind only the beneficial microbes that will help protect the casing and/or bulk substrate.


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Offlineinked4life
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Re: Multi spore syringe and questions on monokaryotic mycelium [Re: insanemike]
    #21966499 - 07/19/15 06:20 PM (8 years, 6 months ago)

yeah i plan to stick to pf tek. im wondering if i should start my new spores on pda til i isolate a good strong mycelium or just innoc brf cakes and roll with it.


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Invisibleinsanemike

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Re: Multi spore syringe and questions on monokaryotic mycelium [Re: inked4life]
    #21966514 - 07/19/15 06:23 PM (8 years, 6 months ago)

If you want a good clone, take a pin from a cluster off one of your cakes and put it on agar. Make a transfer or two to make sure its clean and then expand onto some grain jars. That's what I would do.


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