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InvisiblerelsseS
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Am I killing my spores? Most jars are duds, everytime.
    #22015678 - 07/29/15 03:43 PM (8 years, 7 months ago)

Hey guys. Quick question...  When I inoculate, I flame sterilize the needle tip in between every jar. Once the needle is RED hot, it only takes about 2 seconds for the color to return back to normal metal, and I proceed to inject the spores into the holes. I've gotten jars to grow plenty of times and never have a contamination, but a lot of the time the jars I just duds. I blamed it on bad rye, bad spores, bad sterile technique, etc, but by now I do everything perfect, everytime.

I do a perfect rye grain prep (I rinse and clean grains vigorously until the water is crystal clear. I soak them for 12-20 hours, depending on room temp, and rinse and clean them vigorously again since the soak-water turns nasty. When the ryes are clean as a whistle, I simmer the grains until they're perfectly plump and proceed to air-dry them until the grains are cracker-dry.
Into the jars they go, and they are PC'ed.

I wrap the shit out of my jars in aluminum foil and yet, no matter how hard I try, the polyfil always becomes wet after removing the foil at inoculation time. I've read this is normal and okay, but isn't that interfering with GE and inviting contam?

They are inoculated in a 2x6, 8' empty and freshly sterilized room. Everything is scrubbed down with hospital-grade 99.9% wipes and lysol-blasted. I then shower, and do the inoculation clean and natural.

They are kept at 85 degrees exactly. The room just happens to rest at that exact temperature with no upkeep due to the hot summer.


My question is, when the needle trip is red-hot, and the red fades to normal metal color after 2 seconds, is the needle too hot and killing my spores? Since I flame sterilize after EVERY jar, I assume the needle gets pretty hot towards the end of the 18-jar process.

Every time, either EVERY jar from that needle grows myc, or NONE of them do. And my jars never get contaminated even when they sit there for weeks, it's just plain ol' rye grain, with the perfect moisture content in the perfect temperatures.

Like, I'll use 2 or 3 different "strain" multispore needles during the inoculation (which is usually 12 to 18 jars at a time). I use between 1 and 2 CCs of water per jar, equally distributing the water in a ring around the outside of the rye near the glass.

Every time, the needle either doesn't grow shit, or all the jars begin to show nice myc. And right now, my success rate seems to be at a low 25%-0% rate, and it's pissing me off.


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OfflinetaGyo
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Re: Am I killing my spores? Most jars are duds, everytime. [Re: relsseS]
    #22015691 - 07/29/15 03:46 PM (8 years, 7 months ago)

85 is hot, go down to 70-80.

Quote:

taGyo said:
Alochol SANITIZES, flames STERILIZE. Sterilization means NO germs, mold can survive sanitation.
Don't wipe your needle with alcohol after flame sterilization:

Quote:

swatt_haze said:
wiping a needle with alcohol after flame sterilizing is like rubbing shit on your hands after you washed them and eating a burger.





Flame until the needle is red hot. It will turn black first (because you're using a lighter), then slowly turn red.

Flame sterilize between each JAR, not each inoculation hole. .25 cc a hole for 1cc total each BRF jar.
Grains is 1CC as well.

After flame sterilization gently push down on the plunger. It'll shoot out steam until a drop of liquid comes out. Let the drop gently fall and then push your needle through the hole, making sure to angle it TOWARDS THE GLASS. I look down and check to make sure my needle is against the glass sometimes but you can feel it.

If you push and it feels difficult and nothing's coming out gently pull your needle upward while shooting. BE CAREFUL, this will immediately loosen up your needle and if you're pushing too hard you'll shoot way too many CCs into a hole.




Use 1cc per jar.

Don't use multiple strains, all from the same syringe.

Are contams growing?


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InvisiblerelsseS
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Re: Am I killing my spores? Most jars are duds, everytime. [Re: taGyo]
    #22015740 - 07/29/15 03:54 PM (8 years, 7 months ago)

What's wrong with using 1.5cc, or 2cc? And no I don't mix multiple "strains" in the same jar. I mean during inoculation, after using a needle for 8-9 jars, it is empty and I grab another needle to finish off the last 10 jars I have, because I usually do 18 jars at a time.


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Invisiblepiratez
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Re: Am I killing my spores? Most jars are duds, everytime. [Re: taGyo]
    #22015751 - 07/29/15 03:55 PM (8 years, 7 months ago)

Are you letting your jars cool completely after the pc cycle?

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InvisibleDr.Satan
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Re: Am I killing my spores? Most jars are duds, everytime. [Re: relsseS]
    #22015812 - 07/29/15 04:09 PM (8 years, 7 months ago)

Quote:

relsseS said:
What's wrong with using 1.5cc, or 2cc? And no I don't mix multiple "strains" in the same jar. I mean during inoculation, after using a needle for 8-9 jars, it is empty and I grab another needle to finish off the last 10 jars I have, because I usually do 18 jars at a time.




I've always read to only use 1cc because anymore then that and you're injecting too much moisture into your jars which throws off the moisture levels and in turn invites contams to grow. Now this would not explain why your jars are duds though. :shrug:


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OfflinetaGyo
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Re: Am I killing my spores? Most jars are duds, everytime. [Re: piratez]
    #22015821 - 07/29/15 04:11 PM (8 years, 7 months ago)

Quote:

relsseS said:
What's wrong with using 1.5cc, or 2cc? And no I don't mix multiple "strains" in the same jar. I mean during inoculation, after using a needle for 8-9 jars, it is empty and I grab another needle to finish off the last 10 jars I have, because I usually do 18 jars at a time.



Waste of spores in grains.

Quote:

piratez said:
Are you letting your jars cool completely after the pc cycle?



:whathesaid:

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Invisiblejbaby007
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Re: Am I killing my spores? Most jars are duds, everytime. [Re: relsseS]
    #22015898 - 07/29/15 04:29 PM (8 years, 7 months ago)

Using 1.5-2cc of spore solution messes up the water content in your jars. It's better to have your mixture a little on the dry side than too wet because contams can set in much easier if it's too wet. Spores are microscopic and it only takes 2 to germinate so it's kind of a waste of solution. That's why it's recommended to use only 1cc max of spore solution.


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OfflinetaGyo
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Re: Am I killing my spores? Most jars are duds, everytime. [Re: jbaby007]
    #22015901 - 07/29/15 04:30 PM (8 years, 7 months ago)

In grains 2cc is fine. When I pour my LI It's WAY more then 2cc.


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Offlinespacechildo
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Re: Am I killing my spores? Most jars are duds, everytime. [Re: taGyo]
    #22015933 - 07/29/15 04:36 PM (8 years, 7 months ago)

spore syringes are never truly clean, use as little as possible to lower the chance of shooting in a contam spore.

You've been here quite some time OP but you dont buy your spores from 101 or ralph do you?

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Invisiblejbaby007
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Re: Am I killing my spores? Most jars are duds, everytime. [Re: taGyo]
    #22015941 - 07/29/15 04:38 PM (8 years, 7 months ago)

Ahh, I just skimmed over the thread. Didn't see grains, thought it was pf :rofl:


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OfflinePrimalSoup
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Re: Am I killing my spores? Most jars are duds, everytime. [Re: taGyo]
    #22015958 - 07/29/15 04:40 PM (8 years, 7 months ago)

No need to resterilize that poor needle for every jar UNLESS you accidentally touch it to something that's not sterile.  Bring it to red hot and then shoot some out to hear the sizzle and cool it off.  NOW you're sterile back into the syringe.  You will never draw that plunger back at any time, so if you heat it up again, there better be more sizzle.  That needle won't stay hot for more than a few seconds and when you shoot the cold contents out it won't stay hot at all.  Contams ain't gonna jump into the needle in between spurts...

But your cultures may be bulk or dead or who knows what, sounds like, if there's no germination and no contams either after a decent time. :shrug: 

BTW 85 is too hot.  Cubes don't like it but contams do, so you're stacking the deck against yourself.  Take it down to mid 70s and cruise.


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OfflinePsilosoulful

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Re: Am I killing my spores? Most jars are duds, everytime. [Re: relsseS]
    #22015997 - 07/29/15 04:50 PM (8 years, 7 months ago)

Quote:

relsseS said:
and proceed to air-dry them until the grains are cracker-dry.
Into the jars they go, and they are PC'ed.



Make sure you don't over dry your grains, you want them to be properly hydrated inside, but the outside to be dry. Test your grains by putting a few on a piece of toilet paper, wait thirty seconds, if it leaves behind any wet spots, they're too wet and continue to air dry. :thumbup:

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Offlinespacechildo
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Re: Am I killing my spores? Most jars are duds, everytime. [Re: Psilosoulful]
    #22016025 - 07/29/15 04:56 PM (8 years, 7 months ago)

oh man, nice catch dude!

cracker dry isnt right you want them plump and filled with water on the inside,
but somewhat dry to the outside. cracker dry is what you want harvested and dried mushrooms to be!

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Re: Am I killing my spores? Most jars are duds, everytime. [Re: spacechildo]
    #22016097 - 07/29/15 05:12 PM (8 years, 7 months ago)

:whathesaid: It's okay to PC grains that are a bit too wet on the outside, because they will dry up a bit through the GE hole in your lid, but grains that are PC'd too dry, will not regain any moisture.

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Offlinematsc
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Re: Am I killing my spores? Most jars are duds, everytime. [Re: Psilosoulful]
    #22016163 - 07/29/15 05:31 PM (8 years, 7 months ago)

Are these vendor syringes, or did you make them yourself? If done yourself, are you sure there is adequate spores in each one? Its possible (though admittedly unlikely) your solution is just so dilute that you aren't getting any spores into the mix.

The spores are hydrophobic and tend to stick to plastic and the surface of water, so if you are just scraping into water and then drawing up, its possible to "miss" the spores and just pull up pure water.


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InvisibleMad Season
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Re: Am I killing my spores? Most jars are duds, everytime. [Re: relsseS]
    #22017046 - 07/29/15 08:24 PM (8 years, 7 months ago)

Try it on agar? :shrug: it is the shit after all


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InvisiblerelsseS
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Re: Am I killing my spores? Most jars are duds, everytime. [Re: Mad Season]
    #22017111 - 07/29/15 08:36 PM (8 years, 7 months ago)

I let my PC cool down overnight before inoculation.

My grains are hydrated to the MAX, but the outside is bone dry.
The moisture levels in the jars are perfect..

And you fucking know what, I checked my jars and the bottom of a few of them have cobweb. A solid green nasty rye grain and white fuzz growing around it. Gonna toss the jars asap.

Anyway, I don't know what I'm doing. And I've tried several vendors and had the same problem.

I want to get a good LC or agar, but first I want 1 fucking multispore inoculation to grow so I can pick a good fruit to clone! I've only done 1 successful batch ever and it was a total surprise, I didn't have the materials to clone.


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OfflinePsilosoulful

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Re: Am I killing my spores? Most jars are duds, everytime. [Re: relsseS]
    #22017136 - 07/29/15 08:40 PM (8 years, 7 months ago)

Quote:

relsseS said:
I want to get a good LC or agar, but first I want 1 fucking multispore inoculation to grow so I can pick a good fruit to clone! I've only done 1 successful batch ever and it was a total surprise, I didn't have the materials to clone.



I would just get the agar supplies now and do spores to agar to get CLEAN growth first, then transfer to grains in an SAB. Less frustration and better results every time. :thumbup:

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InvisibleMad Season
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Re: Am I killing my spores? Most jars are duds, everytime. [Re: relsseS]
    #22017159 - 07/29/15 08:45 PM (8 years, 7 months ago)

In my opinion and from my developed experience, every project should be started and cleaned up on agar. Including spores. It's the basis for my communal grow. It's also a great habit to get into. It isn't just for clone work. It's literally the starting point for everything. Lcs included. It's the only way to start 100% clean. 100% of the time.


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Edited by Mad Season (07/29/15 08:48 PM)

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OfflinecronicrFacebook
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Re: Am I killing my spores? Most jars are duds, everytime. [Re: Mad Season]
    #22018131 - 07/30/15 01:49 AM (8 years, 7 months ago)

where are they syringes coming from? i ask because i just ordered some lc syringes and they came in the mail PIPING hot


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