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OfflineNightPuma1
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Can I inoculate a petri dish using a colonized rye berry? * 1
    #26653411 - 05/06/20 05:45 PM (3 years, 8 months ago)

Can I take a fully colonized library and drop it onto a petri dish to inoculate it?

I keep trying to do it from syringe but for some reason I'm getting zero growth no matter what. I've used like five syringes and they're all from a trusted vendor.

Normal agar recipe 7.5 grams LME, 10 grams agar. I shake up the syringes, put a drop on the center and get no growth. I have a bunch of grain jars that I knocked up from syringe that probably aren't 100% clean but maybe I could start on the agar from there.

Thanks.


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Invisiblepanne cyanne
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Re: Can I inoculate a petri dish using a colonized rye berry? [Re: NightPuma1]
    #26653414 - 05/06/20 05:47 PM (3 years, 8 months ago)

yes you can.


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OfflineLogicaL ChaosM
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Re: Can I inoculate a petri dish using a colonized rye berry? [Re: NightPuma1]
    #26653527 - 05/06/20 06:36 PM (3 years, 8 months ago)

Yup! As long as its clean spawn, its perfectly fine.


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OfflineNightPuma1
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Re: Can I inoculate a petri dish using a colonized rye berry? [Re: LogicaL Chaos]
    #26653540 - 05/06/20 06:40 PM (3 years, 8 months ago)

Honestly it probably isn't but I've been failing at agar for far too long with syringes and I'm not waiting for some spore prints to arrive so...


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OfflineLogicaL ChaosM
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Re: Can I inoculate a petri dish using a colonized rye berry? [Re: NightPuma1]
    #26653552 - 05/06/20 06:43 PM (3 years, 8 months ago)

How have your syringes failed? Are u using an inoculation loop?


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OfflineMrBovineJoni
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Re: Can I inoculate a petri dish using a colonized rye berry? [Re: NightPuma1]
    #26653565 - 05/06/20 06:48 PM (3 years, 8 months ago)

Quote:

NightPuma1 said:
Honestly it probably isn't but I've been failing at agar for far too long with syringes and I'm not waiting for some spore prints to arrive so...




Well, you can inoculate but be ready that you will have to clean up and transfer on some more agar unless that grain was taken from a sterile jar inside SAB/under flowhood (very unlikely), so you can avoid spores but you can't avoid agar.

I just did exactly that with some Azurescens that one member here kindly sent me (no prints available so he just sent me a small piece of colonised grains/substrate).


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OfflineNightPuma1
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Re: Can I inoculate a petri dish using a colonized rye berry? [Re: LogicaL Chaos]
    #26653584 - 05/06/20 06:53 PM (3 years, 8 months ago)

No I do not use an inoculation loop but the number of plates that I have put a droplet on should at least have some sort of growth of some kind yet none of them do. And I've used about four different syringes from a very reputable source.

Instead of using an inoculation loop to spread it out, I do tilt the petri slightly to let the droplet run a little (but never touch the wall). I mean that has to be better than nothing and I just don't want to risk another contamination vector by sticking and inoculation looping at this time.

Regardless, putting a drop on 40 different plates from four different syringes should result in something.

Procedure: still air box everything wiped down with tons of alcohol (again my problem is no growth not contamination). Put one droplet right in the center of the petri dish, then I hold the dish slightly diagonally to let the drop run a little bit (I don't feel comfortable doing an inoculation loop so this is at least a little bit of spreading it out.

Yes I shake the bejesus out of the syringes before use. I get nothing growing on any of these. I've used probably 4-5 syringes from a reputable source whose syringes have always worked on other mediums.

I'm going nuts here.


Edited by NightPuma1 (05/06/20 06:55 PM)


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OfflineMrMong


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Re: Can I inoculate a petri dish using a colonized rye berry? [Re: NightPuma1] * 2
    #26653722 - 05/06/20 07:36 PM (3 years, 8 months ago)

Recently I nearly went mad because i noced like 20 plates with 2 different syringes and got nothing weeks later. At one point i genuinely thought the syringes had no spores in.

I later had some success when I changed my agar recipe (to 2g ME: 0.8g agar: 100ml water) and incubated them - got germination within a week on most plates. On a few plates i injected a stupid amount of MS solution, like 1 ml and the plate was swimming, after a week in the incubator all the water evaped and its now COVERED in myc. Albeit this method will give increase contamination risk, it got me the mycelium.

I think increasing water content and temp, as well as the amount of spore solution you use will give you a good chance at germination.


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InvisiblefahtsterM
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Re: Can I inoculate a petri dish using a colonized rye berry? [Re: MrMong] * 3
    #26654244 - 05/07/20 01:48 AM (3 years, 8 months ago)

I’d love to see a fully colonized library.. rr is the obvious librarian choice there

Faht


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Offlinepoisoned
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Re: Can I inoculate a petri dish using a colonized rye berry? [Re: fahtster]
    #26654258 - 05/07/20 02:08 AM (3 years, 8 months ago)

When did you inoculate the plates?


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OfflineScandinavian
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Re: Can I inoculate a petri dish using a colonized rye berry? [Re: poisoned]
    #26654310 - 05/07/20 02:54 AM (3 years, 8 months ago)

I did 1ml pr plate, then transfer at the first sign of mycelium growth.
Usually there will be quite a few colonization points, choose the best looking ones and then just toss the MS plate. πŸ‘Œ

Or use a z pattern with an innoculating loop, I dip mine in agar before MS solution. Never had a failed plate.

I did however have 8 plates with LC that didn't colonize at all.


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Offlinemushpunx
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Re: Can I inoculate a petri dish using a colonized rye berry? [Re: Scandinavian] * 2
    #26654328 - 05/07/20 03:23 AM (3 years, 8 months ago)

"
Instead of using an inoculation loop to spread it out, I do tilt the petri slightly to let the droplet run a little (but never touch the wall). I mean that has to be better than nothing and I just don't want to risk another contamination vector by sticking and inoculation looping at this time."

You can get away with this but really you are going to have to learn to open plates and use sterilezed tools to work with agar. The spore solution it'self is going to be the most likely source of contamination, better to swipe it and give you a better chance of identifying clean my to transfer. :shrug:


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Offlinemushpunx
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Re: Can I inoculate a petri dish using a colonized rye berry? [Re: mushpunx] * 2
    #26654333 - 05/07/20 03:26 AM (3 years, 8 months ago)

Thing about putting a colonized grain on agar, if there was any bacteria or mold in your grain spawn (and many cultivators have bacteria in their spawn and don't know it, they just get lower yields than they should, it will transfer along with the grain to the plate.
And may be quite hard to separate.


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InvisibleLadysKnight
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Re: Can I inoculate a petri dish using a colonized rye berry? [Re: mushpunx] * 1
    #26654666 - 05/07/20 08:33 AM (3 years, 8 months ago)

When I have germination problems I make brf cakes, then clone the fruits.


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OfflineLogicaL ChaosM
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Re: Can I inoculate a petri dish using a colonized rye berry? [Re: LadysKnight]
    #26654820 - 05/07/20 10:19 AM (3 years, 8 months ago)

Ive heard BRF agar plates work well. Bodhi did a thread tek on it.


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OfflineOregonic
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Re: Can I inoculate a petri dish using a colonized rye berry? [Re: LogicaL Chaos]
    #26654888 - 05/07/20 11:01 AM (3 years, 8 months ago)

I revived a clone from a... Very... Old jar of grain spawn by putting a grain onto agar. Used sterilized tweezers to grab out a grain. It took a ton of transfers to get a clean/normal looking culture and tbf I haven't grown it out yet, but the culture does look healthy.

Strange you have had such poor luck with spore syringes to agar. It sounds like you're doing everything right, but if the spores will germ on grain then they should certainly germ on agar. Dumb question but you are cooling the needle after flaming the needle before inoculation, right?


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Offlinemushpunx
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Re: Can I inoculate a petri dish using a colonized rye berry? [Re: Oregonic]
    #26655107 - 05/07/20 12:46 PM (3 years, 8 months ago)

It can be tricky to get spores to germinate on agar sometimes. Sometimes adding a little less agar to soften it up a bit can help, or I'll switch from MEA to PDA etc


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Offlineihuntbacon
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Re: Can I inoculate a petri dish using a colonized rye berry? [Re: mushpunx]
    #26969849 - 10/04/20 09:16 PM (3 years, 3 months ago)

i'm about to try this from rye to agar.


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