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InvisibleOeric McKenna
LIFE CAPS


Registered: 06/15/12
Posts: 5,318
Loc: Babylon Flag
No Pour Culture TEK
    #18014923 - 03/26/13 04:37 PM (11 years, 5 days ago)

 

Do you enjoy the ease of injections, but hate the unpredictable nature of cultures??

I like to see what I'm working with (agar) and then not open it again.
 

This is a combination of previously explained methods:

Sfd/injection port lid TEK.
No pour agar TEK.
LC TEK.
   
    The agar is made in baby food jars or other small jars. The lids are doctored per previously outlined methods.
Sterilize the jars in your PC.
Make & sterilize your LC of choice in a jar with the aforementioned lids.

Sterilize agar jars.
    When your fruit is picked, clone tissue sample to agar jar.
As many as you like.  The clean samples are either transferred to more jars or when you are ready, inject your LC solution from a PC'd LC jar (previously made, & cooled) through the port onto the clean mycelium.
  Wait 4 or more days .(not too thick for ease of use*)
With sterile syringe, tilt agar/lc and suck out the contents.
Test it. Expand it.


  Any questions or comments, feel free.

All of this is geared towars 2 concepts:
1) giving away cultures that someone with limited skill will have high success rates using.
  If the culture is clean, very high success at inoculation is easy.
2) The ease of injectables without the guessing
3) not opening ANYTHING after the initial agar cloning. I feel this could be helpful to people without flowhoods or who may have a high contaminant spore count in their location.



Edited by Oeric McKenna (06/27/13 12:03 AM)

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Invisibleb plus

Registered: 10/13/12
Posts: 928
Re: No Pour Culture TEK [Re: Oeric McKenna]
    #18016869 - 03/26/13 10:58 PM (11 years, 5 days ago)

Should have tossed this over in general cultivation. More people could benefit from it:thumbup:

That's a really great Idea. I've been using sterile distilled water, and scrapping the surface of the myc with a the needle before pulling the myc water back into the syringe. So barbaric compared to your no pour LC agar tek.


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:mushroom2:-Reishi Grow-:mushroom2:
:mushroom2:-Grey Oyster Grow-:mushroom2:

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InvisibleOeric McKenna
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Registered: 06/15/12
Posts: 5,318
Loc: Babylon Flag
Re: No Pour Culture TEK [Re: b plus]
    #18017273 - 03/27/13 02:17 AM (11 years, 5 days ago)

I did consider posting there actually. I'm on an edible kick and I figured edible growers might have the precepts down already.  Someone could move it. Mods?


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OfflinePurpleHead


Registered: 01/17/13
Posts: 128
Loc: South Africa
Last seen: 9 years, 5 months
Re: No Pour Culture TEK [Re: Oeric McKenna]
    #18017357 - 03/27/13 03:09 AM (11 years, 5 days ago)

I sort of did the same thing.  Took a couple of baby food jars, and added an injection port to the lid, no filter.  Transferred mycelium from a successful clone to the bottle and let it grow out.  There seemed to be enough air inside the bottle to do this without GE, but if your growth is slow or you have a slow species I do not think it will work.

Injected sterile water through the port, shook the bottle vigorously, extracted mycelium laden water and transferred to a pint jar full of LME liquid culture solution.

Worked well for me.

Thanks for the great tips!

Looking for those pics.....


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I'm a Noob! If I give advice use it at your own discretion, and please correct me if I'm wrong. I learn form my mistakes, and so can you.

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InvisibleOeric McKenna
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Registered: 06/15/12
Posts: 5,318
Loc: Babylon Flag
Re: No Pour Culture TEK [Re: PurpleHead]
    #18017408 - 03/27/13 03:51 AM (11 years, 5 days ago)

I think the uploader has issues. It keeps saying "uploading"..
& yeah, mycelium water is good too. The reason I use malt extract water (sometimes with wood chunk) is that the culture will grow & expand. You're right that you don't really need the breather port but if you do have them, you can use the same jars for culture growth. Growing culture jars exhaust more.


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OfflineDarkman
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Registered: 04/16/11
Posts: 225
Loc: Africa
Last seen: 7 years, 5 months
Re: No Pour Culture TEK [Re: Oeric McKenna]
    #18017659 - 03/27/13 06:40 AM (11 years, 5 days ago)

Interesting technique.  I don't support LC, but since you mentioned this recently
in another thread, my interest has piqued.

I went out and bought three baby bottles and plan on making a very light substrate
only agar (wood).  I will then pour the bottle as a slant and inoculate with a wedge.
Once the slant is grown over, inject sterile water below the level of the slant, shake and wait
(with the bottle standing upright).

If contaminants don't show up on the agar, aspirate and use to inoculate steamed sawdust spawn.
I am combining this with some trials using sawdust spawn bags.

What do you think?

Also, LC being a tricky topic over here at Edibles, could you please state your level of success
(contam rate) with this Tek, i.e you have used it for 14 years with > 10,000 jars (95%),
or you "winged" 5 out of 6 jars in the past 3 months.

Please don't move the thread, I will post my results here (unless of course the whole Shroomery
calls me a damn fool for trying lol:o)

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InvisibleOeric McKenna
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Registered: 06/15/12
Posts: 5,318
Loc: Babylon Flag
Re: No Pour Culture TEK [Re: Darkman]
    #18018256 - 03/27/13 10:24 AM (11 years, 4 days ago)

(My success rate over the last year with has been unbelievably high.  Here's why:
You get a visual before the culture ever begins. After that point, it is never opened again.
Important to remember is the sterile syringe handling procedure. Do not expose a syringe that has the plunger pushed forward to open air. 

Once you test the culture (& it will likely be clean) you can expand it freely.
I feel that the bad reputation of liquid cultures is because they often do not start on the grounds of certainty as this method does.  IF YOU INVEST TIME IN UNCERTAIN CULTURES YOU WILL WASTE TIME/MONEY AND SPREAD MOLD AROUND.

Keep in mind that the culture syringes sold by vendors are the dreaded "LCs" haha.

Lastly, I trust that injecting clean culture through an injection port in a grain jar has a far higher success rate than agar to grain inoculation.  No matter how good your flowhood is, you are still opening the jar. I'm also sure the common source of contaminants is the individual doing the work!
  If I told you my success rate you wouldn't believe it but let me say that I am a fan of both liquid innoculant and proper syringe handling procedure.
    So, I understand how someone could be against LCs but I honestly don't partake in grain to grain transfers. Grain so readily contaminates.

  I'm not sure about a baby bottle and how the seal is because I've never used one. Also not sure how they fare in a PC but I wish you luck.
I've also never inoculated sawdust and I'm not sure how it would grow or how you'd disperse it .
I personally use grain as a starter spawn.  I wish you luck with that also.


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Offlinedrake89
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Registered: 06/26/11
Posts: 4,168
Loc: TN
Last seen: 5 years, 27 days
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Re: No Pour Culture TEK [Re: Oeric McKenna]
    #18018720 - 03/27/13 12:03 PM (11 years, 4 days ago)

FWIW LC pan. stip. LC stalled out for me on sawdust but took off on PF jars with tef rather than rice flour, as well as P-corn.  Also I had good luck with my first wild clone and LC, chicken of the woods.  This gave me false confidence and all the rest of my LC attempts have failed.  However, I'm ready to re-start now that I have a proper setup (flowhood, autoclave flasks and stirrers, mason jar blender etc...)


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Fiery Fungi (like us on faeboo)

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OfflineDarkman
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Registered: 04/16/11
Posts: 225
Loc: Africa
Last seen: 7 years, 5 months
Re: No Pour Culture TEK [Re: drake89]
    #18023113 - 03/28/13 09:54 AM (11 years, 3 days ago)

Thanks for the tip, it's  been a long time since I did LC
and I had forgotten about the back of the syringe.

I got the baby bottle idea from here, I think it
might be 6T/agar's Tek.

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InvisibleOeric McKenna
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Registered: 06/15/12
Posts: 5,318
Loc: Babylon Flag
Re: No Pour Culture TEK [Re: Darkman]
    #18024017 - 03/28/13 01:47 PM (11 years, 3 days ago)

I think a lot of people don't realize that about syringes.


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OfflineHumbled
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Re: No Pour Culture TEK [Re: Oeric McKenna]
    #18024888 - 03/28/13 05:07 PM (11 years, 3 days ago)

I like the idea Mckenna! I'll have to do this next time I mess with agar. It seems that this would ensure the purity of LC's.

I like using those injection port lids. So many uses:awesome:


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OfflinePurpleHead


Registered: 01/17/13
Posts: 128
Loc: South Africa
Last seen: 9 years, 5 months
Re: No Pour Culture TEK [Re: Oeric McKenna]
    #18026924 - 03/29/13 02:02 AM (11 years, 3 days ago)

Quote:

Oeric McKenna said:
I think a lot of people don't realize that about syringes.




I sure never did. But it makes sense when you think about it as the area inside the syringe is exposed then. Go figure.

Please note that when i speak about babayfood jars i talk about tiny glass jars with metallids and not the bottles which they drink milk from.


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I'm a Noob! If I give advice use it at your own discretion, and please correct me if I'm wrong. I learn form my mistakes, and so can you.

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Invisiblephalcon005

Registered: 12/21/05
Posts: 217
Re: No Pour Culture TEK [Re: Oeric McKenna]
    #18057693 - 04/04/13 01:34 PM (10 years, 11 months ago)

Quote:

Oeric McKenna said:
inject your LC solution through the port onto the clean mycelium.
  Wait 4 days.
With sterile syringe, tilt agar/lc and suck out the contents.





Do you agitate the LC solution at all in that four day period?  I love LC but always had trouble with clumping mycelium, which would clog a syringe.  Methods of cutting it were just enough of a hassle to drop LC completely at the time.  I was likely using oyster or reishi spawn,when I was having clumping issues.  What strains have you used?

Beyond the clumping issue, these teks make LC look dreamy, will have to fire some up.

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InvisibleOeric McKenna
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Registered: 06/15/12
Posts: 5,318
Loc: Babylon Flag
Re: No Pour Culture TEK [Re: phalcon005]
    #18057928 - 04/04/13 02:22 PM (10 years, 11 months ago)

Yes , agitation is good.
I've tried Oyster, shiitake, shaggy mane,lions mane, king stropharia, maitake, cubensis, zapotecorum, & caerulescens.
Oyster seemed the thickest. Shiitake the thinnest.  If a chunk clogs the syringe just push it forward a hair or shake it off.  Having a good sized needle helps a lot.

  Thanks for checking out the method dude


--------------------


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love is everything
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OfflineMyco_tyson
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Registered: 10/27/20
Posts: 7
Last seen: 2 years, 10 months
Re: No Pour Culture TEK [Re: Oeric McKenna]
    #27143219 - 01/12/21 06:05 AM (3 years, 2 months ago)

Hi, thanks, great tek.. I'm having trouble with the turning it to an lc part.. The liquid goes very cloudy and the top of the agar goes mushy, to the point where I can hardly see if there's mycelium growing.
Is this normal or is my agar too soft or something?.. Doesn't look contaminated before adding liquid.

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Offlinetroy_white42
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Registered: 12/04/20
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Re: No Pour Culture TEK [Re: Myco_tyson]
    #27143244 - 01/12/21 06:46 AM (3 years, 2 months ago)

Questions:
Did you flame sterilize and disinfect the area/gloves?
Xfer in front of flow hood?
Was the LC cloudy before xfer?  How many days before xfer?
How many days since xfer?
Pic of your jar?

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OfflineSHROOMSISAY01M
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Registered: 01/22/17
Posts: 3,851
Loc: Virginia, USA
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Re: No Pour Culture TEK [Re: troy_white42]
    #27144228 - 01/12/21 05:07 PM (3 years, 2 months ago)

7-year-old thread. You probably won't get an answer.

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OfflineMyco_tyson
Stranger
Registered: 10/27/20
Posts: 7
Last seen: 2 years, 10 months
Re: No Pour Culture TEK [Re: troy_white42]
    #27145350 - 01/13/21 08:38 AM (3 years, 2 months ago)

Hey, thanks..
No flow hood, and tbh my sab success rate sucks. But out of many agar jars a few looked clean once grown out,but maybe they weren't?
I used an lc mix sterilised in a bottle, then sucked/injected into my agar jar.. All with self healing ports & filters so if It was contams could only have been from my agar xfer, but this grew out nice and looked clean... Its like the sterile lc mix has mixed with the agar and formed something between the 2, like a sludge.. Which now has mycelium growing on the top on 1 jar where I only injected 10ml of liquid.
I thought maybe this was a normal reaction when doing this, or that maybe i prepared my agar (lme/agar/water 20g/20g/1000g) wrong and it was too soft..
But maybe its just contaminated?..



Apologies for the bad pics

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Offlinetroy_white42
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Re: No Pour Culture TEK [Re: Myco_tyson]
    #27145384 - 01/13/21 08:55 AM (3 years, 2 months ago)

So, you only need a drop or two from a syringe onto your agar or it gets all mushy and gross.  I try suck up a tiny piece of mycelium and not too much of the actual water so I don't have to shoot much on.  Your not suppose to make LC from your agar jar.  You grow the agar and when you get a nice clean wedge you know it's not contam.  Then you make another jar with a self heal port and a filter and make a LC mix out of water, lme, and honey or whatever your rcp is. 

Without a hood or sab you'll be hard pressed to cut a small piece of the agar out and drop it into your jar.  Id suggest shooting about 1cc of sterile water into your grown agar jar swirling it around and sucking it hack up.  Then inject that 1cc into your LC jar.  Or if you can reach just try to suck up a little mycelium without water.  Just make sure the agar plate is fully grown and/or there's zero contam growing otherwise when you shoot your water in it will also mix with the cobtam and defeat the whole process.

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OfflineMyco_tyson
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Registered: 10/27/20
Posts: 7
Last seen: 2 years, 10 months
Re: No Pour Culture TEK [Re: troy_white42]
    #27145664 - 01/13/21 11:11 AM (3 years, 2 months ago)

OK, yeah i get you.. I'll try like that..
Just sounded like OP was making the lc on top of the agar.

"inject your LC solution from a PC'd LC jar (previously made, & cooled) through the port onto the clean mycelium.
  Wait 4 or more days .(not too thick for ease of use*)
With sterile syringe, tilt agar/lc and suck out the contents."

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