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Invisiblegenesis128
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MS Inoculation to Agar and DIRECT Transfer to Grain
    #26613380 - 04/19/20 04:46 PM (3 years, 9 months ago)

Can I get some of your guys experiences regarding this?

I want to inoculate an agar dish with a ms print. And without transferring to any more plates, I'd like to transfer that only plate to grains. Would the myc be too weak or would it be ok? Looking for a lot of experiences and advice.

...I would base my decision to transfer further based on all of your info.

Thank you


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Offlinemimir
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: genesis128]
    #26613415 - 04/19/20 05:05 PM (3 years, 9 months ago)

The whole point of going through agar is the ability of remove any contaminants and propagate clean mycelium indefinitely. Doing a single plate doesn't really accomplish any of the two, if that's how you plan to do it it's not worth the trouble.


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Invisiblegenesis128
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: mimir]
    #26613421 - 04/19/20 05:09 PM (3 years, 9 months ago)

I’m not worried about contaminants. If it contams then it contams. Hopefully it doesnt But the reason why im going the agar route is because i dont want to make syringes at the moment. For reasons...


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Offlinemimir
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: genesis128] * 1
    #26613424 - 04/19/20 05:12 PM (3 years, 9 months ago)

Well if a contaminant shows up on agar, are you still going to inoculate jars knowing you'll have to throw them out? If you have spore prints and can't make syringes for some reason I guess you don't have many alternatives, but you'll see contaminants way before the plate is fully colonized anyway


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InvisibleRoger Clemency
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: genesis128]
    #26613433 - 04/19/20 05:17 PM (3 years, 9 months ago)

It’s fine to do it that way, it’s just more risky than transferring a time or two to make sure your clean.


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Invisiblegenesis128
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: mimir]
    #26613435 - 04/19/20 05:18 PM (3 years, 9 months ago)

Naw i wouldnt use that plate if it contams for sure. Ew


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Invisiblegenesis128
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: Roger Clemency]
    #26613438 - 04/19/20 05:19 PM (3 years, 9 months ago)

Thanks for the feedback so far you guys. Please keep them coming


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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: genesis128]
    #26613446 - 04/19/20 05:23 PM (3 years, 9 months ago)

Myc is myc. If it’s clean there’s no problem.

If you got a clean germ plate and used it that would be the same as using a spore syringe.


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Invisiblegenesis128
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: A.k.a]
    #26613466 - 04/19/20 05:33 PM (3 years, 9 months ago)

Yeah... i thought so. Just not sure why the first plate usually looks so tomentose and weird unless transferred to another plate...?


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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: genesis128] * 1
    #26613468 - 04/19/20 05:34 PM (3 years, 9 months ago)

Cuz it’s tons of spores/genetics growing on top of each other. Once it gets thinned out over a couple transfers it looks more uniform.


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Invisiblegenesis128
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: A.k.a]
    #26613478 - 04/19/20 05:40 PM (3 years, 9 months ago)

Oooo nice


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Offlinehuangdeez
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: genesis128]
    #26613516 - 04/19/20 06:05 PM (3 years, 9 months ago)

I say just try it and post about what happens, cuz for newbies like me its good info.


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InvisibleHtaeh
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: huangdeez]
    #26613570 - 04/19/20 06:35 PM (3 years, 9 months ago)

I put a couple chunks out of a ms inoc'd plate which didn't appear to have any contams into two separate grain jars tonight (have other transfer plates growing healthily to fall back on). I wanted to see how it went also thus doing it. Not much to add but I spose I'll post back how it looks in a couple days or so haha.


--------------------
"To write is to wage war" ~ Voltaire
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Invisiblegenesis128
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: Htaeh]
    #26613582 - 04/19/20 06:43 PM (3 years, 9 months ago)

Yes!!  :mushroom2:


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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: genesis128]
    #26613620 - 04/19/20 07:01 PM (3 years, 9 months ago)

So you want fast mushrooms? MS to a LC and roll the dice. It sounds like that is what you are gunning for. LC seems quicker if you don't care about cleaning up the genetic variance and isolating away from contams.


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Invisiblealaskappalachian
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: Sockadin]
    #26613632 - 04/19/20 07:06 PM (3 years, 9 months ago)

Quote:

Sockadin said:
So you want fast mushrooms? MS to a LC and roll the dice. It sounds like that is what you are gunning for. LC seems quicker if you don't care about cleaning up the genetic variance and isolating away from contams.




Ain't it? :awesomenod: That's how I pumped out spawn for kiddie pools 20 years ago lol.  When it worked it was just glorious.  When it didn't there was no lemonade to be made...


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Offlinepoisoned
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: alaskappalachian] * 1
    #26614356 - 04/20/20 05:03 AM (3 years, 9 months ago)

I did that now when I was starting to grow again. Inoculated the plate with spores, did some transfers and threw that plate to grain.

I don't agree it's the same as going MS to grain. You get to see if there's any obvious contamination. Spores also germinate faster on agar. And if you introduced any contam spores when nnocing the jar, myc still has a headstart.


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InvisibleHtaeh
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: poisoned] * 1
    #26614376 - 04/20/20 05:17 AM (3 years, 9 months ago)

Exactly, you at least get the opportunity to observe the myc before hand. Yes there are a lot of genetics growing upon each other so its possible for contams to be buried. I had some plates which had no food colouring in and the agar went pretty clear, when I put a light beneath them I could see on two of the ms plates an area of a contam, as an attempt to garner more experience I transferred from those two plates onto others and they are now looking healthy; I had two other ms plates which appeared to have no contams, the plates were deliberately made very thin and as they appeared clean I decided to practice grain prep and A2G transfer from one and the other one as a transfer to new food coloured plates which are looking brilliant, my best plates to date. So should my A2G transfer succumb to contamination I'll have first hand experience to avoid doing it in the future from a ms plate. And as stated have plenty of healthy plates to fall back on.


--------------------
"To write is to wage war" ~ Voltaire
"The Drug Medical System cannot bear examination. To explain it would be to destroy it.." ~ R.T. Trall M.D.
"A single lie destroys a whole reputation for integrity." ~ Baltasar Gracian (1601-1658), Spanish philosopher 
"We'll know our disinformation program is complete when everything the US public believes is false."  ~  William J. Casey, 1981
"Fear always works to influence the population, when the population is uneducated."
"The secret of freedom lies in educating people, the secret of tyranny is in keeping them ignorant." ~  Maximilien Robespierre
"Knowledge makes a man unfit to be a slave."  ~  Frederick Douglass


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Invisiblegenesis128
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: Htaeh]
    #26625274 - 04/24/20 05:27 PM (3 years, 9 months ago)

Bump... hteah... how are those ms jar coming along??


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Invisiblemushhead
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: genesis128]
    #26625351 - 04/24/20 06:11 PM (3 years, 9 months ago)

Hey genesis128 I've done a few things with MS directly to grain. Yes it does work and you can check out my thread about it to see, however I strongly suggest you make a few transfers to build strong myc.

https://www.shroomery.org/forums/showflat.php/Number/26604440


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Invisiblegenesis128
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: mushhead]
    #26625483 - 04/24/20 07:18 PM (3 years, 9 months ago)

Yeah! That's a nice looking jar. Thanks for the update!


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InvisibleHtaeh
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: genesis128]
    #26626241 - 04/25/20 04:45 AM (3 years, 8 months ago)

Well I made two jars, one is a 1litre tub and the other a small band and lid jar. The intention was to use the ms nocc'd agar to see how it went (using very thin plates and the cleanest looking ones). Happy to report both jars have continued healthy growth. Will post a pic in a couple days or update if disaster strikes, feeling optimistic! :grin:


--------------------
"To write is to wage war" ~ Voltaire
"The Drug Medical System cannot bear examination. To explain it would be to destroy it.." ~ R.T. Trall M.D.
"A single lie destroys a whole reputation for integrity." ~ Baltasar Gracian (1601-1658), Spanish philosopher 
"We'll know our disinformation program is complete when everything the US public believes is false."  ~  William J. Casey, 1981
"Fear always works to influence the population, when the population is uneducated."
"The secret of freedom lies in educating people, the secret of tyranny is in keeping them ignorant." ~  Maximilien Robespierre
"Knowledge makes a man unfit to be a slave."  ~  Frederick Douglass


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OfflineLogicaL ChaosM
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: genesis128]
    #26626313 - 04/25/20 06:07 AM (3 years, 8 months ago)

As long as the culture on your agar is clean, drop a chunk in a grain jar! Thats how its done.


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InvisibleHtaeh
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: LogicaL Chaos]
    #26647424 - 05/04/20 04:16 AM (3 years, 8 months ago)

Spawned the ms agar grain to a shoebox this morning. The grain smelt of fresh mush so hoping for good results. Other clean transferred grain jars are colonising well. Very happy with the surface conditions of the shoebox, had to manually get the coir to field capacity but oh well. Did 1:1, I planned on CVG but didn't bother with verm :shrug:  just coir and gyp and cased with coir. I then clicked the lid on and put it in a bin bag out the way, can anyone tell me whether there is something better to do or if just leaving it in the bin bag for the next 10 days is the best port of call?


--------------------
"To write is to wage war" ~ Voltaire
"The Drug Medical System cannot bear examination. To explain it would be to destroy it.." ~ R.T. Trall M.D.
"A single lie destroys a whole reputation for integrity." ~ Baltasar Gracian (1601-1658), Spanish philosopher 
"We'll know our disinformation program is complete when everything the US public believes is false."  ~  William J. Casey, 1981
"Fear always works to influence the population, when the population is uneducated."
"The secret of freedom lies in educating people, the secret of tyranny is in keeping them ignorant." ~  Maximilien Robespierre
"Knowledge makes a man unfit to be a slave."  ~  Frederick Douglass


Edited by Htaeh (05/04/20 04:52 AM)


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Offlinemimir
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: Htaeh]
    #26647511 - 05/04/20 05:33 AM (3 years, 8 months ago)

Hey! I followed this TEK with pretty good results


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InvisibleHtaeh
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: mimir]
    #26647531 - 05/04/20 05:50 AM (3 years, 8 months ago)

Top guy thanks.


--------------------
"To write is to wage war" ~ Voltaire
"The Drug Medical System cannot bear examination. To explain it would be to destroy it.." ~ R.T. Trall M.D.
"A single lie destroys a whole reputation for integrity." ~ Baltasar Gracian (1601-1658), Spanish philosopher 
"We'll know our disinformation program is complete when everything the US public believes is false."  ~  William J. Casey, 1981
"Fear always works to influence the population, when the population is uneducated."
"The secret of freedom lies in educating people, the secret of tyranny is in keeping them ignorant." ~  Maximilien Robespierre
"Knowledge makes a man unfit to be a slave."  ~  Frederick Douglass


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Offlinemushpunx
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: genesis128]
    #26647658 - 05/04/20 07:50 AM (3 years, 8 months ago)

Quote:

genesis128 said:
Can I get some of your guys experiences regarding this?

I want to inoculate an agar dish with a ms print. And without transferring to any more plates, I'd like to transfer that only plate to grains. Would the myc be too weak or would it be ok? Looking for a lot of experiences and advice.

...I would base my decision to transfer further based on all of your info.

Thank you




You should always transfer at least once.
Why wouldn't you? I transfer as soon as I see clean growth. It would only add two days to your grow.
Prints are never %100 clean , because the mushrooms are grown in open air. When you inoculate, you swipe the spores across the plate in a zig zag pattern. It makes it easier to identify clean myc to transfer after germination.
Contams like bacteria or mold colonies can sometimes be hidden in plates, especially in germination plates. Agar gives us a 2D surface to isolate clean myc from contams. It's the whole point.

Otherwise, you might as well make spore syringes from your print :shrug:


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Invisiblegenesis128
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: mushpunx]
    #26677379 - 05/17/20 06:03 PM (3 years, 8 months ago)

any updates on this project? anyone got shrooms pumped out yet? I just put some agar to a spawn bag and it's colonizing but pretty slowly.... Way too slow. I wonder if my grains are too dry? Can anyone confirm how grains are supposed to look after pressure cooking? Are they supposed to be pretty wet looking or dry? Please help

I can't tell if my culture is slow or my grains aren't proper.


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Invisiblegenesis128
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: genesis128]
    #26677839 - 05/17/20 10:03 PM (3 years, 8 months ago)



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Offlinemushpunx
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: genesis128]
    #26677881 - 05/17/20 10:40 PM (3 years, 8 months ago)

I think it's actually better to prep grains a bit on the dry side.
How big of a wedge did you inoculate with?


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Invisiblegenesis128
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: mushpunx]
    #26677899 - 05/17/20 11:01 PM (3 years, 8 months ago)

Probably about 40 %of the agar dish. Another One was 20% of the agar dish. I want to see if it'll be able to colonize at that tiny size.

Why do you like it dry? For what reason?


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Offlinemushpunx
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: genesis128]
    #26678033 - 05/18/20 01:29 AM (3 years, 8 months ago)

I just think it does better than wet.

Those wedges are fine sizes, how much grain did you put in the spawn bags?

Once a size about the size of my fist is colonized, I break it up and shake the spawn. It usually finishes colonization in a few days after that.


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Invisiblegenesis128
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: mushpunx]
    #26678035 - 05/18/20 01:31 AM (3 years, 8 months ago)

I’d say each bag had 1-3 quarts haha... didn’t really weigh it out. It was extra grain. I made too many. Will report back on how they do.


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InvisibleHtaeh
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: genesis128]
    #26680245 - 05/19/20 03:43 AM (3 years, 8 months ago)

Good morning! A quick update on my germination plate to grain to shoebox. It's day 14 since spawn and WE got pins :biggrin:

All other grain jars are very healthy (much more violent myc in comparison to the germ noc up) but I only really noc grains with agar pins at the moment. I feel its a sure fire way, with little room for error.

To add to what mushpunx said below, dryer grains in my opinion seem to perform better, I've had to experiment with the water content (oats) recently and the only two that contaminated (albeit I know exactly why they did) they were indeed considerably wetter than the ones that perform well and haven't contaminated.

As for colonisation times, as long as it doesn't stall and you continue to notice growth even small amounts of growth its worth letting them continue as you have a lot to gain and nothing to lose. Colonisation times vary, I've got a jar almost ready which was nocc'd ten days ago, one that's about 30% and that was nocc'd the same time so its just about patience really.


--------------------
"To write is to wage war" ~ Voltaire
"The Drug Medical System cannot bear examination. To explain it would be to destroy it.." ~ R.T. Trall M.D.
"A single lie destroys a whole reputation for integrity." ~ Baltasar Gracian (1601-1658), Spanish philosopher 
"We'll know our disinformation program is complete when everything the US public believes is false."  ~  William J. Casey, 1981
"Fear always works to influence the population, when the population is uneducated."
"The secret of freedom lies in educating people, the secret of tyranny is in keeping them ignorant." ~  Maximilien Robespierre
"Knowledge makes a man unfit to be a slave."  ~  Frederick Douglass


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Invisiblegenesis128
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: Htaeh]
    #26681133 - 05/19/20 01:13 PM (3 years, 8 months ago)

Very Interesting Htaeh!
/
By agar pins I'm assuming you wait til your agar pins then put it to grain


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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: genesis128]
    #26681157 - 05/19/20 01:25 PM (3 years, 8 months ago)

Or do you just drop the little pin by itself to grain? curious


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OfflineA.k.aM
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: PMBastian]
    #26681159 - 05/19/20 01:27 PM (3 years, 8 months ago)

You pop the pin off the plate and toss it in the jar.

I’ve got some rusty whyte pins on grain myself.


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OfflinePMBastian
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: A.k.a]
    #26681168 - 05/19/20 01:29 PM (3 years, 8 months ago)

cool, does that colonize quicker than just putting wedges to grain? never had an agar pin before, but i've left my agar plates out this time around in hopes of getting them.


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OfflineA.k.aM
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: PMBastian]
    #26681177 - 05/19/20 01:33 PM (3 years, 8 months ago)

Well.

Cronicr made a thread and his was extremely fast.

Mine have not been but I also didn’t drop the pins until they were close to an inch long. I dropped two at first and it took about four days to see myc starting but really only on the cap and very end. But then I dropped a third younger pin the other day and the whole thing is starting to grow myc after two days.


So I think the key is to get it on grain ASAP.


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InvisibleHtaeh
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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: A.k.a]
    #26681416 - 05/19/20 03:48 PM (3 years, 8 months ago)

Quote:

genesis128 said:
Very Interesting Htaeh!
/
By agar pins I'm assuming you wait til your agar pins then put it to grain




As A.K.a said above, the plate will pin if left long enough and then you transfer said pin to the grain itself.

Quote:

A.k.a said:
Well.

Cronicr made a thread and his was extremely fast.

Mine have not been but I also didn’t drop the pins until they were close to an inch long. I dropped two at first and it took about four days to see myc starting but really only on the cap and very end. But then I dropped a third younger pin the other day and the whole thing is starting to grow myc after two days.


So I think the key is to get it on grain ASAP.





I decided to try three things in three separate Jars to compare.

First was a pin, real small young pin. Second, a more matured pin/mush half way between a knot and breaking the veil. Third, a fully matured sporulating mushroom (on agar of course).

The first small pin, I pretty much lost for about a week, no amount of rolling the jar around could I find it. After day seven I noticed a green spot, thought it was contam and took it downstairs to chuck it out but instead just left it inside a cupboard, long story short that was the pin turning blue not green and now the jar is over 50% colonised with healthy violent myc. Day 11 since nocc'd.

Also day 11 since inoculation the second more mature pin is over 80% colonised, this pin took less time to turn back from mush chitin to mycelium onto the grain. It's currently just recovered from a shake, again, healthy 'violent' myc.

The third, fully matured mush, took even less time to transfer back to myc and onto the grains. Its just four days since inoculation but still less that 10% colonised.

I surmise that somewhere in between the mush life cycle is best for grain transfer. The small young pin took considerably longer to transfer into mycelium and onto the grain compared to the 'half way' second pin. And although the third one is again quicker than the second to transfer back into myc and onto the grain the second still seems quicker and with healthier myc by comparison. Understandably, its still too soon to tell really, given the third one has only been nocc'd four days ago.

Of course this was just one little test, genetics are different etc but they are all AA+.

Above all, it is a very straight forward and quick inoculation of grain. Using sterile tweezers its an almost fool proof and sure fire way of getting clean spawn.


Edited by Htaeh (05/19/20 03:56 PM)


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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: Htaeh]
    #26681458 - 05/19/20 04:15 PM (3 years, 8 months ago)

I did consider using a very old one.

I should probably do it just to see. It looks perfect cuz it’s like four inches long, dropped spores weeks ago and now it’s already growing fuzz all over it.

Thing would probably take to grain immediately.


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Re: MS Inoculation to Agar and DIRECT Transfer to Grain [Re: A.k.a]
    #26681612 - 05/19/20 06:03 PM (3 years, 8 months ago)

I'd say definitely get it in some grain and see.


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