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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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EZ-Port Needleless Connectors for LC use * 1
    #10364498 -

Edit- updated version of the build shown here. This thread explains theory and use.

LC is a useful technique for many home growers.

It's a challenge as well. It tests your ability to produce and handle sterile cultures under the worst conditions. Because of the inherent risk of contamination, many users choose not to try LC, or fail if they do try.

In my opinion, the major risk for contams is handling the jars. Making a lid system that allows for easy access with minimal handling was my design goal. Combined with a magnetic stirrer, it greatly improves chances of success. Even manual stirring, it improves the odds.

Punch or drill a hole in a lid, then put a syringe cap through the hole.
Use a syringe body as a handle. You may need to tap it with a hammer.



If you are doing Grain LC, punch two larger holes also:


Apply RTV silicone around the syringe cap. Fill in the injector port if you have one. Use a piece of cardboard to smooth the RTV and remove excess.


Add a polyfil puff to your Grain LC lid. Or Tyvek, whatever.


IMPORTANT:
Once the RTV sets, trim the bottom of the syringe cap.
You now have an EZ_Port Needless Connector!


Using your favorite LC recipe, make a jar:
DO NOT use more than 1 cup in a quart jar. Having the jar too full is another contamination hazard. You can sterilize a breather syringe if you want, just wrap it in foil. But a quart jar with only one cup of LC will have a good bit of air in it already. Use a marble or stirbar as appropriate.


Shoot through the foil.
Wipe the top of the foil with alcohol, then press the needle all the way down until the hub holds the foil in place. Inject. Peel back foil, remove foil and syringe with smooth motion, attach breather port. Or use micropore tape.


For Grain LC jars, use only half the normal amount of grain or less. You'll shoot through the injection port, and leave the foil in place until the end, so use two or three layers, and lock it down with the jar ring.


There's a million ways to grow LC. When yours is done, you harvest it in 60 mL syringes like this:


Fill your syringes and cap them, then discard teh unused LC. The jar has been compromised, you now have a higher risk of contams. It's not worth trying to save a penny's worth of LC. Test each syringe before shooting up dozens of jars with it.


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You make it all possible. Doesn't it feel good?

Edited by Doc_T (11/20/09 12:43 PM)

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Re: EZ-Port Needleless Connectors for LC use [Re: Doc_T]
    #10364571 -

Nice TEK! thanx

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Re: EZ-Port Needleless Connectors for LC use [Re: 79towncar]
    #10364721 -

Nice lid.






I like using my needle though, it helps me to target areas of mycelium so I get a concentrated mycelium content.


since your using the stirrer bar, you will have it thouougly mixed beforehand.


I would say also a good idea to just make enough for one large syringee, I believe that it would be compromised afterwards, since the foil will be punctured and you will have an exposed hole once you take off your syringe.


I would be careful with polyfill on grain LC jars. If you inject water and you shake it's going to really get that shit nasty. I would stick with some tape.


Also, why do you have 3 holes? Wouldn't you just need one for the syringe, and one for gas exchange?


careful with that RTV , you may need to take some sandpaper to the cap  or whatnot to make sure that it sticks well.


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c        l        i      c        k        m        e

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Re: EZ-Port Needleless Connectors for LC use [Re: aspore]
    #10364790 -

> why do you have 3 holes? Wouldn't you just need one for the syringe, and one for gas exchange?
You could do it either way. With 3, the needleless port remains untouched until you are ready to withdraw the LC. Or you could have that, plus an injection port to use the needle. Lots of options.

You're right about the RTV. I try to wipe up the excess with a tissue, but I still make a bit of mess.


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You make it all possible. Doesn't it feel good?

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Re: EZ-Port Needleless Connectors for LC use [Re: Doc_T]
    #10631459 -

Hey Doc T just made some lids using your tech going to try them out on my grain lc,(I am making my master lc from a multi spore grain lc), and my 2nd gen lc's.

Do you use the same lids with the same set up for your master lcs?


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Fine line between fishing, and standing on the shore with a pole in your hand.

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Re: EZ-Port Needleless Connectors for LC use [Re: River77]
    #10631495 -

For making LC? I like the one with polyfil or tyvek, and an injection port as well as a withdrawal port. For storing, I just use syringes.


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You make it all possible. Doesn't it feel good?

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Re: EZ-Port Needleless Connectors for LC use [Re: Doc_T]
    #10631542 -

You store your master LC's in syringes? 
And you use the selfhealing port for injection and then the needless port for extraction?

Another question. How do you sterilize the tip of the syringe before you connect it to the port on the jar for extraction?


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Fine line between fishing, and standing on the shore with a pole in your hand.

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Re: EZ-Port Needleless Connectors for LC use [Re: River77]
    #10631560 -

Answer to first two is 'yes'.

The syringe tip is sterile. You can sterilize the whole thing in the PC with foil over the end. The draw up your LC, cap it, and store it.


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You make it all possible. Doesn't it feel good?

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Re: EZ-Port Needleless Connectors for LC use [Re: Doc_T]
    #10631602 -

I like this a lot! Thanks a bunch. Going to knock up some rye with a Pan Cyan and a KSSS tonight that are headed for LCs.

What about when you attach the needle after you have filled the syringe with LC. What do you do for sterilization there?


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Fine line between fishing, and standing on the shore with a pole in your hand.

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Re: EZ-Port Needleless Connectors for LC use [Re: River77] * 1
    #10631823 -

The port is sterile under a little piece of foil locked under the band.
If you work in a glovebox, you don't need to worry about the port too much. In open air, I cover it with an alcohol pad. And I try to minimize the movements- that takes some planning and thinking. Have the cap ready, and the next syringe too.


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You make it all possible. Doesn't it feel good?

Edited by Doc_T (07/06/09 06:56 PM)

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Re: EZ-Port Needleless Connectors for LC use [Re: Doc_T]
    #10631861 -

Many Mahalos. I think that these lids are going to rock!


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Fine line between fishing, and standing on the shore with a pole in your hand.

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Re: EZ-Port Needleless Connectors for LC use [Re: River77]
    #10653023 -

just use a jar half full of liquid and a solid dome lid, open it insidea  glovebox and you can save 10x the time :wink: if you dont have a glovebox tis the time to make one!


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German Kahuna said:
Fuck you, you fucking retard! I really tried to remain calm but now you managed to make me pop a vein, you ASSHOLE! @me :rofl2:

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Re: EZ-Port Needleless Connectors for LC use [Re: bumber]
    #10654518 -

Anyone is free to use any method they like- but let me point out that nobody who handles sterile solutions professionally would "just open a jar in a glovebox".
If that amount of risk is ok with you, that's great. But using a needleless connector will help with keeping things sterile.


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You make it all possible. Doesn't it feel good?

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Re: EZ-Port Needleless Connectors for LC use [Re: Doc_T]
    #10654622 -

I used this method myself for my first LC clone job.  One thing extra that I added was a self healing injection port made of RTV silicone on the oposite site.  I must admit, I'm still a bit confused about

Quote:
Shoot through the foil.
Wipe the top of the foil with alcohol, then press the needle all the way down until the hub holds the foil in place. Inject. Peel back foil, remove foil and syringe with smooth motion, attach breather port. Or use micropore tape


I was/am under the impression that a needle doesn't come in to play with this setup, other than inoculating.  Are you inoculating through the same hole that you'll be drawing out through?

I basically inoculated through the other RTV port and left the EZ port covered with foil the whole time.  I hope I didn't screw it up.

Of course this doesn't leave any room for gas exchange... is that necessary with LC, should I remove the foil and cover the EZ port with micropore tape?


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So heavy I fell through the earth...


Edited by Lostkeys (07/10/09 07:51 PM)

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Re: EZ-Port Needleless Connectors for LC use [Re: Lostkeys]
    #10654926 -

Shoot through the foil unless you make a separate injection port.
By doing it the way you have, you are keeping the withdrawal port totally virginal until the LC is done. But it's a more complicated build.
So there you go- but I like to do it the same way yo do, with polyfil and an injection port, as well as the withdrawal port. For Grain LC jars too.

Edit- depending on how much airspace, may or may not need to provide GE. But you can put a sterilized filter syringe on there, as shown.

Edit again- I said you used polyfil. :facepalm: If you decide the syringes are clunky, you can use polyfil in the lid. Either way. Or just leave room- a cup of LC in a quart jar should colonize fine.



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You make it all possible. Doesn't it feel good?

Edited by Doc_T (07/10/09 09:15 PM)

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Re: EZ-Port Needleless Connectors for LC use [Re: Doc_T]
    #10654949 -

umm virginal.....

    I made to jars with a GT and they were taking off, I checked them today and some ants were drinking my LC!  Little bastards. going to need to put the foil on tighter around the extraction port next time. Nice tek though the ants didn't get a Pan Cyan and I just used it  knock up some qt rye. It was really smooth. My props again to this tek Doc T :hairmetal:


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Fine line between fishing, and standing on the shore with a pole in your hand.

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Re: EZ-Port Needleless Connectors for LC use [Re: Doc_T]
    #10654993 -

Doc_T said:
Anyone is free to use any method they like- but let me point out that nobody who handles sterile solutions professionally would "just open a jar in a glovebox".
If that amount of risk is ok with you, that's great. But using a needleless connector will help with keeping things sterile.


ive never heard of someone doing spawn professionally use polyfil either, they mostly use microfiber filter discs...but yea Im not a pro and my glovebox is sterile enough to do agar transfers in, in a slow and steady fasion...and the room is hepa filtered, no contams yet.

just saying this takes time if you have to make more than 2-3 jars with these lids, time that I would spend doing other things :wink:


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German Kahuna said:
Fuck you, you fucking retard! I really tried to remain calm but now you managed to make me pop a vein, you ASSHOLE! @me :rofl2:

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Re: EZ-Port Needleless Connectors for LC use [Re: bumber]
    #10655151 -

Yeah, the first few took a while to do. Now it's pretty easy.
I've made some with Tyvek, that's more 'pro', maybe. :lol:


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You make it all possible. Doesn't it feel good?

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Re: EZ-Port Needleless Connectors for LC use [Re: Doc_T]
    #13626411 -

Nice write up Doc!
I will have to try these ut someday,
Anyone interested in LC lids should check these out too:grin:

https://www.shroomery.org/forums/showflat.php/Number/13621387/fpart/1/vc/1


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Re: EZ-Port Needleless Connectors for LC use [Re: Barakanaten]
    #13626432 -

As stated in that thread, those are not PCable.


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You make it all possible. Doesn't it feel good?

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