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Innoculating LC (Liquid Culture)


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#1 hungryhippo

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Posted 26 February 2008 - 07:55 AM

My FOAF is going to innoculate a jar of Hippies Dextrose water tomorrow and has a few questions. He is worried because he lost 5 PF jars to contams out of 24, and he doesn't want to contam his liquid culture. He doesnt have a glovebox and was going to use regular precautions like lysol, gloves, mask, flame the needle, then wipe the needle with alcohol and inject 2cc multispore. Now, after this LC has grown mycelium, he's worried about repeated needles being stuck into the LC, since he lost 5/25, that is 20% failure rate. Would it be better if he stuck the first needle in the injection port of the LC and left the same needle in while switching out syringes-kind of using the needle like an IV to draw up each syringe, or better to use a new needle with each syringe which you actually have to stick the LC more that way.

He also has one extra syringe of mulitspore and one more jar of Hippies dextrose water and wonders if they would be best stored separately, or should he innoculate the second LC jar. He will have so much LC from the first jar he just wonders the best way to store the rest and keep it good as long as possible. thank you

#2 antimatt3r

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Posted 26 February 2008 - 08:47 AM

personally i make my LC in the 1/2 pint jars, i mix the karo with water, pressure cook or boil, then i simply put a piece of tape over the hole, whenever I need to innoculate, im sure that my syringes have been pressure cooked and stored in a sterile environment (baggie) I remove the syringe from the baggie, wipe the needle part with alcohol soaked paper towel, then insert the needle thru the tape and draw up as much LC as needed, as long as im using the same strain, if I need more LC after ive innoculated however many jars i just wipe the needle again with alcohol and suck more up. upon completion i simply put a new piece of tape over the hole.......... havent had any problems yet!
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#3 onediadem

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Posted 26 February 2008 - 04:41 PM

Try cutting a piece of a brand new sponge about 3/4 inch square and a little longer than the length of your needle. Sterilize the syringe and needle with the sponge on it. After being sterilized, wet the sponge with alcohol so its damp. You have to have some sort of port on your LC jar to push the sponge and needle against to draw up the LC. Leave the sponge on to inoculate your PF style jars, the verm barrier will catch any alcohol, but this shouldn't be an issue if you haven't saturated the sponge. Grain jars need to have the sponge pulled back before inoculating since there is no verm barrier. This works really well if you have had a lot of contam issues and no glove box. Your jars have to have holes drilled in them however if you do not have a glove box..

#4 hungryhippo

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Posted 26 February 2008 - 07:39 PM

Onediadem-that is a veryt interesting tek! I have never read or heard of it? Have you or anybody else used it with great success?

what about the storage of the extra jar and extra syringe of multispoire? Better to innoculate the second jar and store like that ( 2nd jar wont be needed for a LONG time with the frist jar of LC ) or to keep them stored separate and innoculate the LC jar further down the road when it is needed?

#5 onediadem

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Posted 26 February 2008 - 08:30 PM

I use it when I'm just too damn lazy to load up the glove box. Works great. It's not really a tek, just a form of laziness on my part. It has never failed me tho. If you plan on using the LC within 6 months, go ahead and knock it up. Otherwise, I would keep them separate until you do need it. I have had LC's last over a year, but it's better to err on the side of caution.

#6 hungryhippo

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Posted 26 February 2008 - 11:06 PM

Try cutting a piece of a brand new sponge about 3/4 inch square and a little longer than the length of your needle. Sterilize the syringe and needle with the sponge on it. After being sterilized, wet the sponge with alcohol so its damp. You have to have some sort of port on your LC jar to push the sponge and needle against to draw up the LC. Leave the sponge on to inoculate your PF style jars, the verm barrier will catch any alcohol, but this shouldn't be an issue if you haven't saturated the sponge. Grain jars need to have the sponge pulled back before inoculating since there is no verm barrier. This works really well if you have had a lot of contam issues and no glove box. Your jars have to have holes drilled in them however if you do not have a glove box..


So I just want to clarify, because I am trying to picture this in my head. My FOAF can have this sheath made from a sponge he keeps over the needle thoughout the ENTIRE injection process? I mean he withdraws the LC with this needle that is encased in an alcohol soaked sponge, then for shooting PF jars, he keeps this same sponge over the needle and then does NOT flame in between jars?

#7 onediadem

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Posted 26 February 2008 - 11:20 PM

Yep, thats your ticket hon. Keep in mind you only want it damp, not saturated. If you get too much alcohol in the sponge, squeeze out the excess. No flaming the needle required.

#8 hungryhippo

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Posted 27 February 2008 - 06:35 AM

Yep, thats your ticket hon. Keep in mind you only want it damp, not saturated. If you get too much alcohol in the sponge, squeeze out the excess. No flaming the needle required.

well that is an awesome idea1 Thank you! I wonder why this idea isn't in the archives or something???

#9 Sloppy Joe

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Posted 15 March 2008 - 01:35 AM

i think hippie has a thread around here calling it the sponge-o-matic. something to that effect.

#10 Hippie3

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Posted 15 March 2008 - 01:46 AM

close you are.
:bow:
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