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Offlinerew736
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Agar to Syringe??
    #18426938 - 06/16/13 10:10 AM (10 years, 10 months ago)

I have a wedge. How would I turn that into a syringe for an LC??


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OfflineJeff
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Re: Agar to Syringe?? [Re: rew736]
    #18426979 - 06/16/13 10:23 AM (10 years, 10 months ago)



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OfflineBloodKil
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Re: Agar to Syringe?? [Re: Jeff]
    #18441819 - 06/19/13 11:01 AM (10 years, 10 months ago)

I'd make sure everything is sterilized and steps were taken inside a gb/by flow hood...


Wedge+water (not distilled, but I'd use purified Imo) goes into sanatized (or sterilized if possible) blender/magic bullet/etc... 

Blend

Take back to sterile area (wipe down)

Suck into sanatized (or sealed) syringe. (Make sure you have a needle guaged to fit the chunky texture)

Flame sterilize needle

Inject

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Invisible36fuckin5
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Re: Agar to Syringe?? [Re: BloodKil]
    #18442187 - 06/19/13 12:34 PM (10 years, 10 months ago)

Just crack the lid of the LC in a flowhood or SAB and drop the wedge in with a sterile scalpel.

But the real question is why do you want to go backward from agar to LC? Either start a new plate or drop it in grain and G2G.


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Invisiblefortheloveofnature
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Re: Agar to Syringe?? [Re: 36fuckin5]
    #18448056 - 06/20/13 04:36 PM (10 years, 10 months ago)

The way I do it is follow the no pour agar tek with half pints but I put a injection port in the lid . When the agar is fully colonized and pure white I get a few syringes of sterilized water and squirt the mycelium a few times till I get a decent amount of mycelium. Than suck the water back up and bam you got yourself some LC . Not the best way but a lot easier if you don't got a flowhood and are working in a SAB.


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InvisibleNecroMyce
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Re: Agar to Syringe?? [Re: 36fuckin5]
    #18454691 - 06/22/13 01:08 AM (10 years, 10 months ago)

Quote:

36fuckin5 said:
Just crack the lid of the LC in a flowhood or SAB and drop the wedge in with a sterile scalpel.

But the real question is why do you want to go backward from agar to LC? Either start a new plate or drop it in grain and G2G.




I was wondering this earlier to. Probably the same reason i would want to. To knock up BRF. But id assume if you were gonna use a wedge to inoculate you wouldn't have to add that extra little layer of vermiculite as a contam barrier.

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OfflinePussyFart
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Re: Agar to Syringe?? [Re: NecroMyce]
    #18454852 - 06/22/13 02:37 AM (10 years, 10 months ago)

If you wanted to knock up a BRF cake with an agar wedge, use a lid with a filtered GE of some kind, and no, no need for a dry verm layer.

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OfflineCyber
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Re: Agar to Syringe?? [Re: PussyFart]
    #18468666 - 06/25/13 05:21 AM (10 years, 10 months ago)

When I make syringes from Agar Culture I do the following

I do it all under the flow hood, I never had good luck with a glove box for agar work.
Take a syringe of sterile water flame sterilize the needle and inject 2cc of water into the petri dish on top of the growth.
Scrape the mycelia growth loose into the drop of water then suck it back up into the syringe. You don't need much to start
Flame sterilize the needle again and inject the strands into an LC.
Once it has colonized the LC I use it to make syringed to inject my spawning media.

You can go directly from the petri-syringe to a BRF cake and skip the LC. I have done this many times and it works fine.

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OfflineJcoonan
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Re: Agar to Syringe?? [Re: Cyber]
    #18488886 - 06/29/13 04:27 PM (10 years, 10 months ago)

Came here to say just that. This is a technique used to culture bacteria, it does work.


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Offlinematsc
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Re: Agar to Syringe?? [Re: Cyber]
    #18489290 - 06/29/13 06:06 PM (10 years, 10 months ago)

I use essentially the same method for inoculation. I grow my samples on small agar plates. When they are as colonized as I want them I add a bit of sterile water, and using flame sterilized tweezers I scrape and disturb the mycelia to get it into solution. With a bit of practice you can avoid getting much agar into the mix, which has a nasty habit of clogging the syringe. (I use 18G 1.5" needles, for reference)

Once everything is in the water I suck it into a fresh syringe and cap it off. I then go straight from this water-mycelia syringe to grain jars. Ive never felt the need to use an LC intermediate. I suppose if wanted, you could do so to amplify the solution and turn the one syringe into many, but I just make more plates if I want to go that route.

This trick has worked fine for everything but ABM, which seems to have a personal grudge against me and refuses to grow at more than a snails pace regardless of what media I toss at it.


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