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OfflineSaBr
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Print --> Syringe --> Innoc --> Nada
    #25348296 - 07/26/18 04:41 PM (2 years, 9 months ago)

I've been successful growing BRF cakes multiple times, all using syringes purchased from sponsors. From those grows, I've begun making spore prints, and subsequently, syringes. I've inoculated using my own syringes now, twice, and both times, no colonization at all. No contam, just no colonization.

What does this suggest? I know my inoculation method is sound, so something is going wrong in my print or syringe making.

Any obvious fail points in either of those processes? Again, no contam, just no life...

Thanks in advance!


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OfflineRombino
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Re: Print --> Syringe --> Innoc --> Nada [Re: SaBr]
    #25348553 - 07/26/18 07:16 PM (2 years, 9 months ago)

Hmmm, good question. I to have made a few syringes from prints I've made from my grows but mine worked. 2 eventually contamed and 2 didn't. How did you transfer print into the syringe? Did you scrape the spores off? Maybe damaged them? I hear they are delicate.... Just a thought...


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Offlinekayo
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Re: Print --> Syringe --> Innoc --> Nada [Re: Rombino]
    #25348602 - 07/26/18 07:47 PM (2 years, 9 months ago)

Thats weird. Could you have killed them somehow? Kept them in a hot car or something? :shrug:


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OfflineSaBr
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Re: Print --> Syringe --> Innoc --> Nada [Re: Rombino]
    #25348628 - 07/26/18 08:00 PM (2 years, 9 months ago)

Quote:

Rombino said:
How did you transfer print into the syringe? Did you scrape the spores off? Maybe damaged them? I hear they are delicate.... Just a thought...




Inoculation loop to scrape from the foil in to a sterilized (alcohol) beaker while in my SAB. Then new, sterile syringe into steam sterilized H2O, that water in to beaker with spores, five times or so of 50ccs and stirred up solution, then draw that solution out to five sterile syringes.

Seems right, no?


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OfflineSaBr
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Re: Print --> Syringe --> Innoc --> Nada [Re: kayo]
    #25348629 - 07/26/18 08:02 PM (2 years, 9 months ago)

Quote:

kayo said:
Thats weird. Could you have killed them somehow? Kept them in a hot car or something? :shrug:




This for certain no. Hot car, heat, light, etc, that is. No idea if I could have killed them some other way. Can't imagine what it could have been.

I may have used a uv-c wand to sterilize the foil, or perhaps even the print... but the wand claims to be spore friendly (specific from a mycology site).


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OfflineRombino
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Re: Print --> Syringe --> Innoc --> Nada [Re: SaBr]
    #25348670 - 07/26/18 08:18 PM (2 years, 9 months ago)

Was all alcohol gone from beaker? Maybe residue from alcohol?:shrug:


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OfflineLtLurker
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Re: Print --> Syringe --> Innoc --> Nada [Re: Rombino]
    #25348839 - 07/26/18 09:48 PM (2 years, 9 months ago)

It's been over a week, enough time to expect to see some life? Alcohol or heat would be my guesses. Heat includes water that's still hot from sterilizing, syringe tip still too hot from flaming, and injecting your jars too hot from the pc.

There's not many other things that would cause zero germination of anything, good or bad, that I'm aware of.

IDK about the uv light thingy. Dunno anyone that used one.


Late thought. Did you use plenty of spores? If you can see specks in there it's enough. I'm thinkin if you made solution for 5 syringes at once, but used only a little scrape of spores, then your syringes may be really diluted.


Edited by LtLurker (07/26/18 09:50 PM)


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OfflineSaBr
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Re: Print --> Syringe --> Innoc --> Nada [Re: LtLurker]
    #25348966 - 07/26/18 10:57 PM (2 years, 9 months ago)

Quote:

LtLurker said:
Late thought. Did you use plenty of spores? If you can see specks in there it's enough. I'm thinkin if you made solution for 5 syringes at once, but used only a little scrape of spores, then your syringes may be really diluted.




Could be alcohol, I suppose, but I dried the beaker pretty thoroughly.

I used a full spore print, so that shouldn't be the issue. However, on that train of thought... the syringes are 'dark' - that is, clearly not clear, as there is obviously some dark 'matter' in there. However I can't see individual spores (I know they're microscopic) or spore clusters, as like vendor syringes. Could this be an indicator of something?


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OfflineLtLurker
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Re: Print --> Syringe --> Innoc --> Nada [Re: SaBr]
    #25349035 - 07/26/18 11:41 PM (2 years, 9 months ago)

No, if you let them sit a little while they'd settle & clump up like a vendor syringe. Sounds like you did fine making them. Letting the alcohol dry should be plenty. I'm kinda stumped.


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Offlineicetech
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Re: Print --> Syringe --> Innoc --> Nada [Re: LtLurker]
    #25350067 - 07/27/18 11:44 AM (2 years, 9 months ago)

Just curious, why not go spores>agar>brf? and skip the syringe?


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OfflineSaBr
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Re: Print --> Syringe --> Innoc --> Nada [Re: icetech]
    #25350337 - 07/27/18 02:33 PM (2 years, 9 months ago)

Quote:

icetech said:
Just curious, why not go spores>agar>brf? and skip the syringe?




Haven't learned agar yet. On the curriculum after grains/bulk.


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OfflineLtLurker
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Re: Print --> Syringe --> Innoc --> Nada [Re: SaBr]
    #25350363 - 07/27/18 02:53 PM (2 years, 9 months ago)

Agar would be a good way to atleast test if the syringe is viable. Would help narrow down where in your process something is wrong. It's super ez to make and well worth learning if you want to use grains. Even clean prints/syringe can be dicey on straight grains.

I started agar before grains(no pc) and would make lc, li, or drop wedges in brf jars to practice.


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Re: Print --> Syringe --> Innoc --> Nada [Re: SaBr]
    #25350403 - 07/27/18 03:24 PM (2 years, 9 months ago)

Quote:

SaBr said:
Quote:

icetech said:
Just curious, why not go spores>agar>brf? and skip the syringe?




Haven't learned agar yet. On the curriculum after grains/bulk.




Honestly... i wish i had skipped the rest and started witha agar, it takes 3 minutes to prep.. then PC and done.. use pastywhites no pour and bod's BRF agar recipe... syringes can blow a goat.. spores on agar go sooo much faster than a syringe also.


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Offlinekayo
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Re: Print --> Syringe --> Innoc --> Nada [Re: SaBr]
    #25350446 - 07/27/18 03:53 PM (2 years, 9 months ago)

Quote:

SaBr said:
I may have used a uv-c wand to sterilize the foil, or perhaps even the print... but the wand claims to be spore friendly (specific from a mycology site).




I couldnt find anything definitive on UVC and mushroom spores but this would be my guess if you did. That thing is used to sterilize and is even dangerous to humans. If you used a whole freshly made print they have to be dead. Either alcohol, uvc, or some other hazard, what else can it be? Sounds like you know how to prepare the jars. So unless someone switched out your flour with talcum powder it should germinate. How long were they exposed, assuming you did sterilize the prints?

I did see a site selling one with "Sterilize spore prints while making your own spore syringes" in the description but damn ... Wish I had a wand, this calls for an experiment!


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OfflineSaBr
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Re: Print --> Syringe --> Innoc --> Nada [Re: icetech]
    #25350663 - 07/27/18 05:59 PM (2 years, 9 months ago)

Quote:

Honestly... i wish i had skipped the rest and started witha agar, it takes 3 minutes to prep.. then PC and done.. use pastywhites no pour and bod's BRF agar recipe... syringes can blow a goat.. spores on agar go sooo much faster than a syringe also.




Good feedback. I I follow the two recipes listed, where do I learn what to do? Are there some simple training TEKs out there?


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Offlineicetech
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Re: Print --> Syringe --> Innoc --> Nada [Re: SaBr] * 1
    #25350845 - 07/27/18 07:43 PM (2 years, 9 months ago)

yup. just do a search.. i usually search google for the bod's brf one.. i have  hard time with the search here.. other is pastywhites no pour agar..  he uses plastic ziplock tubs..i  actually perfect little half pint jars with screw lids.. i have reused them like 10 times now.. i hate throwing things away. 

Actually don't search for the agar.. it's simple
400ml water (1 3/4 cup)
9g agar
6g brf

Get water to low boil.. dump in agar and brf, add food coloring of choice (i like dark colors as i can see the myc against it better. you don't have to use any if you don't want)

stir it up.. once it's disolved put about 1/8th inch thick in the bottom of the jars (thinner is better. i did really thick once and it SUCKED for transferring)  then PC it... it takes minutes.. so easy.

Here is the link to pasty's thread on the no pour..

Pasty's no pour.

I suck at growing shrooms but my agar is always perfect:) doing it this way has always worked great for me.


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Edited by icetech (07/27/18 07:44 PM)


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OfflineSaBr
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Re: Print --> Syringe --> Innoc --> Nada [Re: icetech]
    #25354827 - 07/29/18 11:00 PM (2 years, 9 months ago)

Good feedback, thanks. I've read through Bods and Pastywhites teks. I get the process of making and inoculating agar. I get the "tiger flip" (I think it as called) of agar to grain. The place I'm coming up short is between the two. What does one look for on agar? When does one cut slices (if chosen route)? What to discard?

It's the actual agar time that I'm a little unclear on. And direction is appreciated.


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OfflineLtLurker
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Re: Print --> Syringe --> Innoc --> Nada [Re: SaBr]
    #25354871 - 07/29/18 11:28 PM (2 years, 9 months ago)

In the main tek list at the top of the forum is a link to a bunch of info and pics for agar.


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Offlineicetech
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Re: Print --> Syringe --> Innoc --> Nada [Re: LtLurker]
    #25355390 - 07/30/18 10:22 AM (2 years, 9 months ago)

Quote:

SaBr said:
Good feedback, thanks. I've read through Bods and Pastywhites teks. I get the process of making and inoculating agar. I get the "tiger flip" (I think it as called) of agar to grain. The place I'm coming up short is between the two. What does one look for on agar? When does one cut slices (if chosen route)? What to discard?

It's the actual agar time that I'm a little unclear on. And direction is appreciated.




There are 100's of threads on agar with pics.. but basically once you make it and PC it.. you can either put spores one it or even use a syringe, the advantage is that you can watch the mycelium grow, it tends to grow in a few patters, which means you can see if you have clean growth or contaminants..

In doing it this way you know you have clean myc going into the jar, and every step you can take to make sure you are working clean is a better chance of success. Also once you get agar rolling you can transfer to new agar and even keep it in the fridge (my last ones sat in the fridge for 6 months and were still fine) It's just all around a massive time saver.

I don't tiger drop myself.. i use a xacto blade wiped with iso and then heated with a torch and cut small wedges and drop them, i can do like 5 jars from 1 small agar.


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Mushrooms, Mycology and Psychedelics >> Getting Started

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