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Offlinek1ng87
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how resistance in mycelium to infection?
    #13017977 - 08/07/10 04:21 PM (14 years, 5 months ago)

how resistant is mycelium to infection. I did some LC and inoculated but am not sure how sterile I was.

Also, I can't find any videos or guidance on how to inoculate w/ mycelium from LC.

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Offlinesepsis
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Re: how resistance in mycelium to infection? [Re: k1ng87]
    #13018032 - 08/07/10 04:35 PM (14 years, 5 months ago)

Please give more information on what you did and how you did it.

Sterilized cakes and grain should be inoculated in a glove box.

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OfflineMycjunky
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Re: how resistance in mycelium to infection? [Re: sepsis]
    #13018047 - 08/07/10 04:40 PM (14 years, 5 months ago)

Yea there's so many different ways you could do it it would help us if you could give us a step by step. You don't always have to inoculate in a glovebox if you use self healing injection ports and a sponge with alcohol but I'm guessing that's not what you did.

It's not the mycelium that you should be worried about being resistant to infection the LC itself has nutes in it that aren't colonized by mycelium and could provide a place for contams to grow, as well as the grain your inoculating is very susceptible to contams before it's colonized.

I'd suggest not even bothering with LC's because it can be very difficult for beginners to get a clean LC, and then nearly impossible to tell if an LC is clean until you noc up jars. Grain lc is the way to go and you can tell from the start if it's contamed.

Edited by Mycjunky (08/07/10 04:41 PM)

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InvisibleDoc_T
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Re: how resistance in mycelium to infection? [Re: k1ng87]
    #13018061 - 08/07/10 04:44 PM (14 years, 5 months ago)

Quote:

k1ng87 said:
how resistant is mycelium to infection.



Not very.

Quote:

k1ng87 said:
I did some LC and inoculated but am not sure how sterile I was.



Be sure. Be sure you are sterile, especially with LC work.


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

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Offlinek1ng87
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Re: how resistance in mycelium to infection? [Re: Doc_T]
    #13018170 - 08/07/10 05:13 PM (14 years, 5 months ago)

yeah...my issue was that i could only get 1inch 28guage syringes from the local pharmacy which weren't helpful at all.

I had my LC in a 1pint jar and the mycelium was floating in the middle. I didnt have injection points in teh LC jars, didn't use a glove box either. I did try my best to sterilize my work area and bleach sprayed the entire room heavily.

since the syringe wasn't working...I ended up trying to use a spoon to scope the mycelium out of the jar and put in the cakes....wow, what a mistake...

I still have one untouched jar of LC left...I was planning on buying some syringes right now online for my one jar of LC i have left....

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OfflineMycjunky
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Re: how resistance in mycelium to infection? [Re: k1ng87]
    #13018634 - 08/07/10 07:10 PM (14 years, 5 months ago)

If you buy syringes do not inject the spores into a LC. spore-liquid culture typically is not a very good method, go with a grain lc.

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OfflinehamloafM
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Re: how resistance in mycelium to infection? [Re: k1ng87]
    #13018777 - 08/07/10 07:50 PM (14 years, 5 months ago)

Quote:

yeah...my issue was that i could only get 1inch 28guage syringes from the local pharmacy which weren't helpful at all.



:pm: 
Quote:

I didnt have injection points in teh LC jars, didn't use a glove box either. I did try my best to sterilize my work area and bleach sprayed the entire room heavily.



Open air will pass with agar, not so much with an LC because it is liquid and once a contaminate mold spore touches it, the spore becomes absorbed by the liquid and infects the whole medium.  LC's must be inoculated in a still air glove box at least, best, in front of a flowhood.
Quote:

.I was planning on buying some syringes right now online for my one jar of LC i have left..



Do that, build you a glove box though, to inoculate the LC in and keep the air in the glove box still while you are inoculating the LC.  Only use 1cc to inoculate your LC with.  With the other 9cc's, do BRF cakes, get fruits, print the fruits, and propagate your grow.  What kind of LC recipe are you using?
HL


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Offlinetokerten
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Re: how resistance in mycelium to infection? [Re: Doc_T]
    #13018935 - 08/07/10 08:30 PM (14 years, 5 months ago)

Quote:

Doc_T said:
Quote:

k1ng87 said:
how resistant is mycelium to infection.



Not very.

Quote:

k1ng87 said:
I did some LC and inoculated but am not sure how sterile I was.



Be sure. Be sure you are sterile, especially with LC work.




doc t by no means am questioning your authority,tbh i even look up
to you as a member but i think its kind of reckless to say that.

to the op mcyjunky hit the nail on the head its uncolonized nute
source that will be prone to contaminates,
some people use more spores to increase the chances of the mycelium geting
a foot hold before contaminates but you dont really get all the
added benefits of lc over spores such as having a smaller genetic variance


--------------------
Please, don't believe everything They tell you

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OfflineTheManWithTheHat
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Re: how resistance in mycelium to infection? [Re: tokerten]
    #13018995 - 08/07/10 08:45 PM (14 years, 5 months ago)

Quote:

tokerten said:
Quote:

Doc_T said:
Quote:

k1ng87 said:
how resistant is mycelium to infection.



Not very.

Quote:

k1ng87 said:
I did some LC and inoculated but am not sure how sterile I was.



Be sure. Be sure you are sterile, especially with LC work.



some people use more spores to increase the chances of the mycelium geting
a foot hold before contaminates but you dont really get all the
added benefits of lc over spores such as having a smaller genetic variance



Works to an extent with substrates and agar, but not with LC.
Also, using more spores also increases your chances of getting contamination. If there is 2 infectious microbes in your syringe, and you use 1 mL of solution, there's only a 2/10 chance of contamination. However, if you use all 10 mL, your chance of contamination increases significantly because now both microbes will be in your solution with free access to nutrients. With the scale that microorganisms grow on, you can be assured that the number will be >> than 2.

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Offlinek1ng87
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Re: how resistance in mycelium to infection? [Re: TheManWithTheHat]
    #13019141 - 08/07/10 09:19 PM (14 years, 5 months ago)

aiight...i think i'm gonna scratch the LC method....I only used 2ccs out of the 12 for LC...I also inoculated two cakes with the spore syringe directly with 1cc into self-healing injection sites (two). I also made 4 air holes but covered them with breathable surgical tape.

Man...I planned on inoculating 4 with mycelium and 2 with spore...I did everything correctly except for the damn syringes!....


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Offlinek1ng87
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Re: how resistance in mycelium to infection? [Re: k1ng87]
    #13022921 - 08/08/10 06:24 PM (14 years, 5 months ago)

what should I do with my other jar of LC and what kind of syringes should I get?

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OfflineTheManWithTheHat
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Re: how resistance in mycelium to infection? [Re: k1ng87]
    #13023055 - 08/08/10 06:47 PM (14 years, 5 months ago)

Quote:

k1ng87 said:
what should I do with my other jar of LC and what kind of syringes should I get?



Inoculate only one jar as a tester for your LC or make a new LC from scratch but make sure you use sterile technique. You'll want any sterile syringes with probably either a 18 or 16 gauge needle. You can order those from vendors/sponsors on the site here.

If you're going more for a qualitative approach than a quantitative approach (e.g. it's your first time) you might as well just make an LC from your spore syringe IF, and only IF, you're sure the syringe contains clean spores that aren't contaminated. SO, if you ordered your syringe from a vendor, you should be fine. But, if you made it, it might not be.

Not the best way to go around doing it, but it'll preserve your precious quantity of spores. Use only about a mL of spore solution though.

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OfflineTheManWithTheHat
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Re: how resistance in mycelium to infection? [Re: TheManWithTheHat]
    #13023063 - 08/08/10 06:48 PM (14 years, 5 months ago)

Most importantly though, you should UTFSE man :tongue:

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InvisibleM11
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Re: how resistance in mycelium to infection? [Re: TheManWithTheHat]
    #13023160 - 08/08/10 07:05 PM (14 years, 5 months ago)

Grain LCs are bomb-diggity.  I highly recommend.  :thumbup:


--------------------
Of course, then there's the problem of eating vermiculite. On the bright side, it makes your poop and teeth glitter.  Just pretend it's christmas.
-RR

Those mushrooms are fine.  Your friend is a pussy.
-RR

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