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OfflineNightPuma1
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Can't inoculate agar from syringe
    #26693365 - 05/24/20 06:27 PM (3 years, 8 months ago)

So I make agar petri dishes and I know they are good and sterile because whenever I drop an inoculated rye berry on them I get good growth. HOWEVER, thus far I have not been able to get any growth coming from a syringe (I do one drop on the center and never see any growth of any kind). All my syringes are from reputable sources and I have tried several of them but I just don't seem to be able to go from syringe to agar.

This might sound stupid but - does anyone go from PF tek to Agar? I'm having such bad luck going straight from syringe that I was thinking I might just inoculate a cake and drop a little piece of cake onto agar.

Thanks!


Edited by NightPuma1 (05/24/20 06:28 PM)


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OfflineMH5109
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Re: Can't inoculate agar from syringe [Re: NightPuma1] * 1
    #26693379 - 05/24/20 06:35 PM (3 years, 8 months ago)

How long are you waiting sometimes spores take their sweet time germinating. Iv had some take a month. Before I seen growth.


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OfflineRapjack
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Re: Can't inoculate agar from syringe [Re: MH5109]
    #26693391 - 05/24/20 06:41 PM (3 years, 8 months ago)

I've only had that happen once, it was because the syringe didn't have many spores and they clumped. I shook it thoroughly and it worked the next attempt.

My other theory would be that food grade agar sometimes has preservatives. If that's the case myc would be able to colonize it but spores won't germinate.


Edited by Rapjack (05/24/20 06:43 PM)


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InvisibleMateja
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Re: Can't inoculate agar from syringe [Re: MH5109]
    #26693404 - 05/24/20 06:45 PM (3 years, 8 months ago)

Quote:

NightPuma1 said:
So I make agar petri dishes and I know they are good and sterile because whenever I drop an inoculated rye berry on them I get good growth. HOWEVER, thus far I have not been able to get any growth coming from a syringe (I do one drop on the center and never see any growth of any kind). All my syringes are from reputable sources and I have tried several of them but I just don't seem to be able to go from syringe to agar.

This might sound stupid but - does anyone go from PF tek to Agar? I'm having such bad luck going straight from syringe that I was thinking I might just inoculate a cake and drop a little piece of cake onto agar.

Thanks!



I just inoculated 8 plates with 4 drops each of Pan Cyan spore solution, that's 32 inoculation spots and I'll be happy to see one or two germination spots in a week :smile:


Getting germination from spore solution>agar will vary in how effective it is based on the quality and density of spores. My advice would be to place 3-4 drops on each plate as to now waste too many plates unnecessary, cause all you need is a tiny germination to get the culture going. Gl :thumbup:


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Edited by Mateja (05/24/20 06:46 PM)


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OfflineNightPuma1
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Re: Can't inoculate agar from syringe [Re: MH5109]
    #26693552 - 05/24/20 07:43 PM (3 years, 8 months ago)

Honestly after about 3 weeks I was trashing them so I should probably wait. Good call.


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OfflineNightPuma1
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Re: Can't inoculate agar from syringe [Re: Mateja]
    #26693555 - 05/24/20 07:44 PM (3 years, 8 months ago)

I will start doing more drops. Thanks.


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OfflineMH5109
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Re: Can't inoculate agar from syringe [Re: NightPuma1] * 1
    #26693570 - 05/24/20 07:51 PM (3 years, 8 months ago)

Yea shake the syringes up a good amount and put a few drops on your plate. What is your agar mixture iv personally found that the softer agar works better for germination.


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Offlinejcm4620
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Re: Can't inoculate agar from syringe [Re: MH5109]
    #26693589 - 05/24/20 07:59 PM (3 years, 8 months ago)

what i will do when i have a hard time getting my pans to germinate on plates is il make some small jars of rye nerries and knock them up with the spores. then once i get germination on the grain il pluck several kernals to plates and then clean up and isolate from there. works great every time👍


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OnlineLogicaL ChaosM
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Re: Can't inoculate agar from syringe [Re: MH5109] * 1
    #26693590 - 05/24/20 07:59 PM (3 years, 8 months ago)

Quote:

MH5109 said:
Yea shake the syringes up a good amount and put a few drops on your plate. What is your agar mixture iv personally found that the softer agar works better for germination.




Excellent advice. Sometimes syringes have very low sperm spore count so you have to really shake that syringe up :thumbup:


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Re: Can't inoculate agar from syringe [Re: LogicaL Chaos] * 3
    #26693623 - 05/24/20 08:12 PM (3 years, 8 months ago)

That's why I always shake my balls up real well before I inoculate the missus


Edited by Apples in Mono (05/24/20 08:54 PM)


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OfflineNightPuma1
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Re: Can't inoculate agar from syringe [Re: Apples in Mono]
    #26693627 - 05/24/20 08:13 PM (3 years, 8 months ago)

As for my question about using a piece of PF cake?


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OfflineNightPuma1
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Re: Can't inoculate agar from syringe [Re: jcm4620]
    #26693631 - 05/24/20 08:15 PM (3 years, 8 months ago)

Can you give me a very specific rundown of how exactly you do this?

I was sort of thinking of doing this myself but the problem is that I usually am used to putting about 2.5 CC's into a whole grain jar and I don't know how to do it without upsetting the water balance (too wet = contamination).

How do you do it sir?


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OnlineLogicaL ChaosM
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Re: Can't inoculate agar from syringe [Re: Apples in Mono] * 2
    #26693698 - 05/24/20 08:38 PM (3 years, 8 months ago)

Quote:

Apples in Mono said:
That's I always shake my balls up real well before I inoculate the missus




But thats like fanning a SGFC. Unnecessary cause that Motion of the Ocean / Hitting it with the Misses :awehigh:

U can shoot up PF cake jars then crumble onto Grain. Theres a tek on that too (forgot the name again). Its pretty simple.


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OfflineNightPuma1
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Re: Can't inoculate agar from syringe [Re: LogicaL Chaos] * 1
    #26693718 - 05/24/20 08:46 PM (3 years, 8 months ago)

No but can I PF Tek to Agar?


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OfflineMH5109
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Re: Can't inoculate agar from syringe [Re: NightPuma1]
    #26693725 - 05/24/20 08:50 PM (3 years, 8 months ago)

Yea if you want, just don't put a verm layer on top.  Then open it up and grab a little bit with tweezers. You could probably just shoot some water in there and draw it back up with the syringe and run that way too. Id be careful doing the water injection cuz if you have a contam in there you have a contam in the syringe water too.


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OnlineLogicaL ChaosM
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Re: Can't inoculate agar from syringe [Re: NightPuma1]
    #26693732 - 05/24/20 08:53 PM (3 years, 8 months ago)

Quote:

NightPuma1 said:
No but can I PF Tek to Agar?




Yes you can. Would be easy since a lot of mycelium grows between the cake and the dried verm layer.


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InvisibleApples in Mono
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Re: Can't inoculate agar from syringe [Re: LogicaL Chaos]
    #26693749 - 05/24/20 09:02 PM (3 years, 8 months ago)

Quote:

LogicaL Chaos said:
Quote:

Apples in Mono said:
That's I always shake my balls up real well before I inoculate the missus




But thats like fanning a SGFC. Unnecessary cause that Motion of the Ocean / Hitting it with the Misses :awehigh:

U can shoot up PF cake jars then crumble onto Grain. Theres a tek on that too (forgot the name again). Its pretty simple.




I just don't figure those 2-3 pumps is really enough to do the job:shrug:


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Re: Can't inoculate agar from syringe [Re: Apples in Mono]
    #26693794 - 05/24/20 09:24 PM (3 years, 8 months ago)

whats your agar recipe ?  maybe its a bit stiff?  that always gives me trouble when going from syringes


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OfflineNightPuma1
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Re: Can't inoculate agar from syringe [Re: TedsDead]
    #26693828 - 05/24/20 09:46 PM (3 years, 8 months ago)

So I try and do the 500 ml water, 10 gram agar 10 gram LME but the problem is that I got the syrup LME. I weighed it out one time and found that about one teaspoon of syrup LME is close to 10 grams so that's just what I do.

I actually made a post not too long ago asking people for a recipe that involved the syrup LME and didn't get a ton of help.


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OfflineMH5109
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Re: Can't inoculate agar from syringe [Re: NightPuma1] * 1
    #26693834 - 05/24/20 09:50 PM (3 years, 8 months ago)

Cut that agar back to 5g. That may help you out. The plates will still be solid but much softer than your original ones. And the nutes arnt set in stone so if you arnt exact its not the end of the world. Iv grown mycelium on 1.5g lme 500ml h2o just fine.


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Edited by MH5109 (05/24/20 09:53 PM)


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InvisibleThe Fresh PrintsS
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Re: Can't inoculate agar from syringe [Re: MH5109]
    #26693844 - 05/24/20 09:59 PM (3 years, 8 months ago)

I waited almost 2 weeks for ps. Cyan to germ on agar from a syringe.


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OfflinePsyche delics
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Re: Can't inoculate agar from syringe [Re: The Fresh Prints] * 1
    #26693888 - 05/24/20 10:25 PM (3 years, 8 months ago)

Do you send a drop through to cool the needle before actually inoculating it. The heat be killing of the spores. I had this happen once and I'm pretty sure this is why.


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Re: Can't inoculate agar from syringe [Re: Psyche delics] * 1
    #26693917 - 05/24/20 10:47 PM (3 years, 8 months ago)

:whathesaid:

gotta do that


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Offlinejbgtaa
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Re: Can't inoculate agar from syringe [Re: TedsDead]
    #26693933 - 05/24/20 11:08 PM (3 years, 8 months ago)

Yeah you need to make sure the needle is fully cooled before inoculation.

Besides that I’d say just buy prints instead of syringes :shrug: in my experience it’s way easier to get good germination and growth when streaking a print, check out c10s method of streaking spore prints onto agar.


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Re: Can't inoculate agar from syringe [Re: TedsDead]
    #26693963 - 05/24/20 11:29 PM (3 years, 8 months ago)

Flame-sterilizing before going to agar seems counterproductive to me. I would just use Iso Alcohol on the tip and thats it.


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Offlinejbgtaa
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Re: Can't inoculate agar from syringe [Re: LogicaL Chaos] * 1
    #26693980 - 05/24/20 11:40 PM (3 years, 8 months ago)

Honestly if you’ve kept your syringes capped and they were made cleanly you don’t need to do any of it 🤷🏻‍♂️ The point of agar is to separate clean growth from contams, not to have perfect growth on t0

It’s definitely best practice to try and get healthy germination on the first plate tho. That said, we don’t flame sterilize spores from prints. Just the vessel to move them. I stream my plates with pre-sterilized scalpel blades and have great results. Uses way less plates in the long run, not having to transfer away from contams a bunch of times that I introduced from any number of places or movements


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Re: Can't inoculate agar from syringe [Re: LogicaL Chaos] * 1
    #26694002 - 05/25/20 12:21 AM (3 years, 8 months ago)

Quote:

LogicaL Chaos said:
Flame-sterilizing before going to agar seems counterproductive to me. I would just use Iso Alcohol on the tip and thats it.





if its a fresh tip... sounds like they've used it a few times now


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Offlinepoisoned
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Re: Can't inoculate agar from syringe [Re: TedsDead]
    #26694031 - 05/25/20 01:13 AM (3 years, 8 months ago)

If you're doing cakes, you may as well clone some nice looking fruits to agar.


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OfflineNightPuma1
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Re: Can't inoculate agar from syringe [Re: poisoned]
    #26694271 - 05/25/20 06:06 AM (3 years, 8 months ago)

Everyone is writing about flame sterilizing the needle. I do this and let it cool down.

Still haven't gotten any petri growth (including contaminants) 🤷🏼‍♂️


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OfflinePMBastian
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Re: Can't inoculate agar from syringe [Re: NightPuma1]
    #26694304 - 05/25/20 06:34 AM (3 years, 8 months ago)

That happened to me a while ago. A GT syringe I know is good (cause it worked straight to grain) just refused to germinate on agar. Must be 2 months ago now and the plates still show zero growth.

Sadly I don't know why tho, so I don't have any tips for you!


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Re: Can't inoculate agar from syringe [Re: PMBastian]
    #26694330 - 05/25/20 06:55 AM (3 years, 8 months ago)

Can you tell us what agar recipe you are using?


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Offlinepoisoned
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Re: Can't inoculate agar from syringe [Re: NightPuma1]
    #26694350 - 05/25/20 07:12 AM (3 years, 8 months ago)

Quote:

NightPuma1 said:
Everyone is writing about flame sterilizing the needle. I do this and let it cool down.

Still haven't gotten any petri growth (including contaminants) 🤷🏼‍♂️




I usually let a few drops go into the bottom of my SAB before inoculating. Spores die at relatively low temperatures, better cool that needle off for real.


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InvisibleLadysKnight
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Re: Can't inoculate agar from syringe [Re: poisoned]
    #26694386 - 05/25/20 07:41 AM (3 years, 8 months ago)

For some spores that refused to germinate on agar, I've had luck with brf pucks. Then transfer growth to agar.


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Offlinejackrabbitcytoplas
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Re: Can't inoculate agar from syringe [Re: AtomHeart]
    #26694410 - 05/25/20 07:54 AM (3 years, 8 months ago)

I've had no problem germinating on brf/verm and cleaning it up on agar, a single drop in the middle is enough, just make enough of them to have a few good ones.
You can also make shallow brf pucks, wait for them to pin and clone the pins so you are sure you are cleaning a fruiting culture.




Regarding going directly from syringe to agar, search for the word "streaking" in c10's Agar Guide  here https://www.shroomery.org/forums/showflat.php/Number/24330957#24330957


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