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Offlinebirdbrain84
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How do you put the needle on the syringe???
    #11672105 - 12/17/09 10:13 PM (15 years, 1 month ago)

I'm pretty sure I fucked it up again.:bitch:

How do you put the needle on the syringe without contaminating it??
I was doing all this in the glovebox, and the needle comes seperate from the syringe. So when I was putting the needle on the syringe i noticed that When I take off one part of it, the spores are directly exposed to the air.  Wouldnt that be instant contamination?
Then once the needle was on, I flame sterlized it till it turned kind of red.  I stopped though because it made some weird noise like bubbling or cracking..its hard to explain.  Then When I was about to inoculate it the liquid from inside the syringe was dripping out! So I probably contaminated it.  Also I think I accidentally sucked in some air into the syringe while it was in the glovebox.  Does that mean I fucked up the syringe?


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InvisibleShea25
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Re: How do you put the needle on the syringe??? [Re: birdbrain84]
    #11672120 - 12/17/09 10:16 PM (15 years, 1 month ago)

Prob not. Its normal for liquid to come out and to hear nosies when flaming the needle. I used to put my needles on in the open air with zero problems now I use a glove box Im sure your fine

Edited by Shea25 (12/17/09 10:17 PM)

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Offlinebirdbrain84
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Re: How do you put the needle on the syringe??? [Re: Shea25]
    #11672183 - 12/17/09 10:28 PM (15 years, 1 month ago)

I'm really hoping..

But this is my third try and I fucked up the first 2 times so ......

I'll see in a few days if I messed up or not.

If I did contaminate the syringe is there any way I could save it?


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InvisibleShea25
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Re: How do you put the needle on the syringe??? [Re: birdbrain84]
    #11672185 - 12/17/09 10:29 PM (15 years, 1 month ago)

Yea you could do some gar work on it

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Offlineshroom239
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Re: How do you put the needle on the syringe??? [Re: birdbrain84]
    #11672197 - 12/17/09 10:31 PM (15 years, 1 month ago)

I think you will be fine. You said you were using a glove box which would reduce the chances of a contamination. I always check my needles before using them to make sure they are secure. And as far as that sound while sterilizing the needle, I say its normal.

Quote:

Shea25 said:
Yea you could do some agar work on it



Agreed.

Edited by shroom239 (12/17/09 10:32 PM)

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Offlinebirdbrain84
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Re: How do you put the needle on the syringe??? [Re: shroom239]
    #11672217 - 12/17/09 10:35 PM (15 years, 1 month ago)

I dont know how to do agar..
I havent had a successful grow yet :frown:

I dont even really know what agar is....?

I cant even get the PF tek right


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OfflineBrainChemistry
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Re: How do you put the needle on the syringe??? [Re: birdbrain84]
    #11672224 - 12/17/09 10:36 PM (15 years, 1 month ago)

Are you making a syringe from a spore print, or do you have a pre-made spore syringe? If you have a pre-made syringe, it should have already came with a needle attached to it...at least thats all I've ever seen.

If you are making from a spore print, you are going to have to expose  the spores to air no matter what you do.

But if you sterilize your working environment sufficiently well with aerosol sprays, there really is VERY little chance of getting contamination from the air. You should be much more concerned about contamination from surfaces (which you should also have sterilized before making your syringe).

The needle should come pre-sterlized and sealed in a narrow plastic tube. You should be able to attach the needle to the syringe WITHOUT removing the plastic tube. When you suck up your spore solution into your syringe, there is no need to sterilize the needle. Only do that when you are ready to inoculate. You should be able to replace the plastic tube cap over the needle to ensure that your needle stays sterile until you are ready to inoculate.

When you go run a flame under the needle, it is normal to hear crackling because there is going to be a few water droplets left inside of the needle after you sucked up your solution.

And if liquid is dripping out your syringe...thats fine, because the solution inside is still sterile, and that is what you are injecting into your substrate.


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InvisibleShea25
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Re: How do you put the needle on the syringe??? [Re: birdbrain84]
    #11672225 - 12/17/09 10:36 PM (15 years, 1 month ago)


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Offlinemushking
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Re: How do you put the needle on the syringe??? [Re: Shea25]
    #11672241 - 12/17/09 10:40 PM (15 years, 1 month ago)

chill out bro, your fine, if you had to be THAT sterile this hobby wouldnt be very popular.
ignore the contamination nazi's and chill out well working,
working well stressed fucks you up.

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InvisibleShea25
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Re: How do you put the needle on the syringe??? [Re: mushking]
    #11672247 - 12/17/09 10:42 PM (15 years, 1 month ago)

dude were telling him he's most likily fine no one here is being a contam Nazi I think you need to chill :thumbup:

Working unclean and lazy will fuck you up

Edited by Shea25 (12/17/09 10:43 PM)

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Offlineshroom239
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Re: How do you put the needle on the syringe??? [Re: mushking]
    #11672256 - 12/17/09 10:44 PM (15 years, 1 month ago)

Agreed mushking.
I was really tweaked at first.
Now I'm a little looser with my sterilization.
Not to much but not as extreme as I was.
Yet to fail.
:shrug:


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Disclaimer: Any picture or statement I post here is from research I do on my free time, I do not actually partake in this hobby.

Edited by shroom239 (12/17/09 10:45 PM)

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Offlinebirdbrain84
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Re: How do you put the needle on the syringe??? [Re: mushking]
    #11672261 - 12/17/09 10:44 PM (15 years, 1 month ago)

I wouldnt be so worried about it if I hadnt failed my first 2 times.
The  second time I tried, I was SO careful about being clean, and it still contaminated.  It even gave me false hope because the first day it was growing really fast and looked like healthy white myc. but then the next day it was all green.


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InvisibleShea25
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Re: How do you put the needle on the syringe??? [Re: shroom239]
    #11672267 - 12/17/09 10:45 PM (15 years, 1 month ago)

Well its good to get into a full sterile mode right from the start,when you start to do Agar/G2G ect its worth it and you will be happy you leaned and practiced quick and efficient procedures 

Edited by Shea25 (12/17/09 10:47 PM)

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Offlinebirdbrain84
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Re: How do you put the needle on the syringe??? [Re: Shea25]
    #11672276 - 12/17/09 10:47 PM (15 years, 1 month ago)

Quote:

Shea25 said:
Read this


http://www.shroomery.org/forums/showflat.php/Number/11585613/fpart/1




Thats pretty much exactly what I did.  I just did what it said on those mushroom videos from RR


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Offlineshroom239
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Re: How do you put the needle on the syringe??? [Re: Shea25]
    #11672282 - 12/17/09 10:48 PM (15 years, 1 month ago)

Well if you are taking all the steps to be sterile and still failing then something along the way is introducing contamination. Think about how you are doing things, what your sterilizing and how you are sterilizing them. After doing a review of the steps you take make adjustments cause something is going wrong.


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Disclaimer: Any picture or statement I post here is from research I do on my free time, I do not actually partake in this hobby.

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Offlinemushking
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Re: How do you put the needle on the syringe??? [Re: shroom239]
    #11672283 - 12/17/09 10:48 PM (15 years, 1 month ago)

i was speaking in boarder terms, none of you are contamination nazi's as far as i know.

did my post mention anything about unclean environments and laziness?
i think shaking well your trying to inoculate a jar might inhabit someones potential and prosperity.

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Offlinebirdbrain84
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Re: How do you put the needle on the syringe??? [Re: mushking]
    #11672328 - 12/17/09 10:55 PM (15 years, 1 month ago)

I think I know what went wrong the first time (actually a few things went wrong)
But I cant think of what went wrong the second time.
I didnt cover the holes with tape and it looked healthy in the beginning, is it possible that some contam got past the dry verm layer?


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InvisibleShea25
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Re: How do you put the needle on the syringe??? [Re: birdbrain84]
    #11672347 - 12/17/09 10:59 PM (15 years, 1 month ago)

Not likily at all I have done hundreds of cakes and the dry verm barrier has not failed me

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Offlineshroom239
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Re: How do you put the needle on the syringe??? [Re: birdbrain84]
    #11672362 - 12/17/09 11:01 PM (15 years, 1 month ago)

Quote:

birdbrain84 said:
is it possible that some contam got past the dry verm layer?



From what I understand its very unlikely. Maybe if you shook them to get air circulating in the jars something got past. But that would be at a point where it wouldn't seem to likely in my opinion, though don't quote me.


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Disclaimer: Any picture or statement I post here is from research I do on my free time, I do not actually partake in this hobby.

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Offlinebirdbrain84
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Re: How do you put the needle on the syringe??? [Re: Shea25]
    #11672372 - 12/17/09 11:03 PM (15 years, 1 month ago)

Maybe it was a mold that starts out white in the beginning and turns green afterwords thats what someone else said in another post when I asked about it.
I didnt shake the jar I just took it off the shelf I had it on to look at it and then put it back.

But I still cant think of what went wrong.

Also does it matter that I sucked up air into the syringe?


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I don't suffer from insanity, i enjoy every minute of it.

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