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InvisiblePestile

Registered: 05/02/13
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First time agar, azures MS syringe, which media should I use?
    #18784275 - 09/01/13 06:39 AM (10 years, 4 months ago)

I have an azurescens MS syringe, from a sponsor, which is pitch black inside. I've tried several inoculation methods, each time it contams green. This will be my first agar experiment, for this I have 17 glass petri dishes that has been wrapped in foil and PC'ed for 30 min.

So I want to see if I can clean it using agar. Stamets recommends using some of the end-substrate in the recipe for a smaller lag period.


Is this desirable when I want to clean the culture? Or should I stick to the PDA/MEA recipes?


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Offlineblojo02184
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Re: First time agar, azures MS syringe, which media should I use? [Re: Pestile]
    #18784355 - 09/01/13 07:45 AM (10 years, 4 months ago)

Since your doing a wood lover, try saw dust agar.


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InvisiblePestile

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Re: First time agar, azures MS syringe, which media should I use? [Re: blojo02184]
    #18789177 - 09/02/13 11:04 AM (10 years, 4 months ago)

bump


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InvisibleFrankHorrigan
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Re: First time agar, azures MS syringe, which media should I use? [Re: Pestile]
    #18789199 - 09/02/13 11:08 AM (10 years, 4 months ago)

Potato dextrose yeast agar works great for woodlovers I've thrown at it.

Do you have any hardwood chips?

Soak a couple handfuls of chips in 500mL of water.

Take the chips out and adjust the water back to 500mL.

Add 10g of agar, 10g of potato starch, 7g dextrose, and 1g yeast (brewers/nutritional). PC and pour.


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InvisiblePestile

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Re: First time agar, azures MS syringe, which media should I use? [Re: FrankHorrigan]
    #18789253 - 09/02/13 11:16 AM (10 years, 4 months ago)

I have all those things, though, the yeast is baker's yeast.

I'll try this recipe out tonight, thanks FH!


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InvisibleFrankHorrigan
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Re: First time agar, azures MS syringe, which media should I use? [Re: Pestile]
    #18789260 - 09/02/13 11:18 AM (10 years, 4 months ago)

Go to the grocery store and grab some brewers yeast, it's cheap...sometimes they hide it in the organics section.

Brewers yeast is inactive. Bakers yeast is not.


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InvisiblePestile

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Re: First time agar, azures MS syringe, which media should I use? [Re: FrankHorrigan]
    #18789268 - 09/02/13 11:20 AM (10 years, 4 months ago)

Quote:

FrankHorrigan said:
Go to the grocery store and grab some brewers yeast, it's cheap...sometimes they hide it in the organics section.

Brewers yeast is inactive. Bakers yeast is not.





Inactive, got it!


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InvisiblePestile

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Re: First time agar, azures MS syringe, which media should I use? [Re: Pestile]
    #18799151 - 09/04/13 01:06 PM (10 years, 4 months ago)

Can I squirt a couple drops out on a piece of foil and transfer from there to my dishes? I'd like to try to get the least amount of black syringe water onto the dishes.


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InvisibleVirginiaGentleman
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Re: First time agar, azures MS syringe, which media should I use? [Re: Pestile]
    #18799325 - 09/04/13 01:49 PM (10 years, 4 months ago)

Black? The azure spores I've seen were almond/chocolate color


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InvisiblePestile

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Re: First time agar, azures MS syringe, which media should I use? [Re: VirginiaGentleman]
    #18799345 - 09/04/13 01:55 PM (10 years, 4 months ago)

Yeah, the syringe is pitch black. They are from a sponsor, though. :shrug:


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Invisiblemycomattie
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Re: First time agar, azures MS syringe, which media should I use? [Re: Pestile]
    #18799378 - 09/04/13 02:05 PM (10 years, 4 months ago)

The syringe I have is very dark also.


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InvisiblePestile

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Re: First time agar, azures MS syringe, which media should I use? [Re: mycomattie]
    #18799447 - 09/04/13 02:22 PM (10 years, 4 months ago)

Quote:

mycomattie said:
The syringe I have is very dark also.




Have you inoculated anything yet? I'm a little skeptical about this black water.


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Invisiblemycomattie
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Re: First time agar, azures MS syringe, which media should I use? [Re: Pestile]
    #18799495 - 09/04/13 02:37 PM (10 years, 4 months ago)

Quote:

Pestilence said:
Quote:

mycomattie said:
The syringe I have is very dark also.




Have you inoculated anything yet? I'm a little skeptical about this black water.




Yup, and grew great!  A bit on the slow colonization side, but totally legit.  I ordered from a very reputable sponsor too.


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InvisiblePestile

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Re: First time agar, azures MS syringe, which media should I use? [Re: mycomattie]
    #18799620 - 09/04/13 03:04 PM (10 years, 4 months ago)

My agar is cooling the in the PC right now, so I'll soon find out if this one is legit. :smile:


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Invisiblemycomattie
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Re: First time agar, azures MS syringe, which media should I use? [Re: Pestile]
    #18799658 - 09/04/13 03:12 PM (10 years, 4 months ago)

I'm sure you'll be good.

I grew the P azure's on modified (Alder saw dust) PF Tek style jars; then spawned them to an Alder chip based sub; all was going good until I had to move, and I ended up dumping the project.  The Ps. azurescens mycelium was brilliantly white and healthy....

Keep us posted on your progress, I'm dabbling with this species now too.


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InvisiblePestile

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Re: First time agar, azures MS syringe, which media should I use? [Re: mycomattie]
    #18799719 - 09/04/13 03:26 PM (10 years, 4 months ago)

Will do! Thanks. :smile:


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InvisiblePestile

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Re: First time agar, azures MS syringe, which media should I use? [Re: Pestile]
    #18800534 - 09/04/13 06:56 PM (10 years, 4 months ago)

One thing I learned tonight: glass petri dishes is a complete waste of time and energy. :mad2:


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Invisiblemycomattie
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Re: First time agar, azures MS syringe, which media should I use? [Re: Pestile]
    #18800572 - 09/04/13 07:07 PM (10 years, 4 months ago)

Quote:

Pestilence said:
One thing I learned tonight: glass petri dishes is a complete waste of time and energy. :mad2:




Really?  I was just going to order a bunch of glass petri's thinking they're better than plastic in the long run!


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InvisibleFooMan
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Re: First time agar, azures MS syringe, which media should I use? [Re: Pestile]
    #18800613 - 09/04/13 07:18 PM (10 years, 4 months ago)

I know this answer is a little late, but you don't need to add any end substrate into the mix, it just helps the mycelium get preconditioned to consume the substrate. I use standard PDA or MEA for most species without any additions. Either way, the spores should germinate and you'll hopefully be able to isolate the myc away from the contams. Adding the end substrate wouldn't hurt or help in that aspect.

Good luck :thumbup:


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InvisiblePestile

Registered: 05/02/13
Posts: 875
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Re: First time agar, azures MS syringe, which media should I use? [Re: FooMan]
    #18800632 - 09/04/13 07:24 PM (10 years, 4 months ago)

Quote:

mycomattie said:
Quote:

Pestilence said:
One thing I learned tonight: glass petri dishes is a complete waste of time and energy. :mad2:




Really?  I was just going to order a bunch of glass petri's thinking they're better than plastic in the long run!




Slippery shit, rattling and curvy lids, way too much hassle getting them out of the bag and the foil only to finds out they are half wet from the PC. The film wouldn't quite stick to the wet surface so that's also bugging me out. I'm gonna try a couple plastic sleeves, see how that works out.

Quote:

FooMan said:
I know this answer is a little late, but you don't need to add any end substrate into the mix, it just helps the mycelium get preconditioned to consume the substrate. I use standard PDA or MEA for most species without any additions. Either way, the spores should germinate and you'll hopefully be able to isolate the myc away from the contams. Adding the end substrate wouldn't hurt or help in that aspect.

Good luck :thumbup:




Cool, I'll remember that.
Thanks!


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InvisibleFrankHorrigan
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Re: First time agar, azures MS syringe, which media should I use? [Re: Pestile]
    #18801176 - 09/04/13 09:39 PM (10 years, 4 months ago)

Quote:

Pestilence said:
One thing I learned tonight: glass petri dishes is a complete waste of time and energy. :mad2:




Yep :shake:


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InvisibleVirginiaGentleman
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Re: First time agar, azures MS syringe, which media should I use? [Re: FrankHorrigan]
    #18801421 - 09/04/13 10:34 PM (10 years, 4 months ago)

I've been eyeballing the Pyrex ones... Is their use finicky?

I'm using the ball elites, USPS tyvek lids, SAB, with a no-pour Tek. They've been cake work, besides some operator errors.

I'm lazy and short on time, which has kept me from going to legit glass petris


--------------------
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InvisiblePestile

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Re: First time agar, azures MS syringe, which media should I use? [Re: VirginiaGentleman]
    #18802309 - 09/05/13 07:11 AM (10 years, 4 months ago)

Quote:

VirginiaGentleman said:
I've been eyeballing the Pyrex ones... Is their use finicky?

I'm using the ball elites, USPS tyvek lids, SAB, with a no-pour Tek. They've been cake work, besides some operator errors.

I'm lazy and short on time, which has kept me from going to legit glass petris




I would say that if you only plan on using the agar for inoculation, glass will work. But I wouldn't make more than a couple dishes for that type of use.
If you're expecting to make several transfers, plastic petri with 3 compartments would probably work better for that.



But onto my last query, does anyone know if I can squirt out a drop from the syringe on a piece of foil and transfer from there to my dishes? Maybe with a scalpel or inoculation loop or something like that... :smile:


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Invisiblemycomattie
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Re: First time agar, azures MS syringe, which media should I use? [Re: Pestile]
    #18802445 - 09/05/13 08:14 AM (10 years, 4 months ago)

I'm glad I saw this before I spent all the extra $$ on glass petri's!

I can't inoculate agar with a drop of MS from a syringe to save my life!  I end up squirting too much and the plate gets contaminated; I've been pretty successful working from prints though.

Here is something you can try - while you're PC'ing your agar, throw in a 1/2 pint jar with some tin foil squares in it; when it's cooled, just move to your SAB or flow hood; open the jar, remove the foil, and try from there.

Might be the only way to minimize the risk of cantams  (using the foil you're describing)?


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InvisiblePestile

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Re: First time agar, azures MS syringe, which media should I use? [Re: mycomattie]
    #18802484 - 09/05/13 08:36 AM (10 years, 4 months ago)

You're right, there's no reason not to try it out. :smile:
Thanks for pointing that out.


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InvisiblePestile

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Re: First time agar, azures MS syringe, which media should I use? [Re: Pestile]
    #18811769 - 09/07/13 02:20 PM (10 years, 4 months ago)

The syringe is toast. Nothing but little green patches on the agar.
I have some colonized wood chips, can I transfer one of those to a dish?


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InvisibleFrankHorrigan
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Re: First time agar, azures MS syringe, which media should I use? [Re: Pestile]
    #18811837 - 09/07/13 02:42 PM (10 years, 4 months ago)

Yes, you can, just clip small pieces and place them over a few dishes.


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Yes, you can bump my old threads with a question.
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InvisiblePestile

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Re: First time agar, azures MS syringe, which media should I use? [Re: FrankHorrigan]
    #18811912 - 09/07/13 03:08 PM (10 years, 4 months ago)

Thank you, Frank.


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InvisibleVirginiaGentleman
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Re: First time agar, azures MS syringe, which media should I use? [Re: Pestile]
    #18812024 - 09/07/13 03:48 PM (10 years, 4 months ago)

I love it when woodies take to chips, they get so rhizomorphic :durrhurr:


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InvisiblePestile

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Re: First time agar, azures MS syringe, which media should I use? [Re: VirginiaGentleman]
    #18857113 - 09/18/13 08:57 AM (10 years, 4 months ago)

So I transferred some colonized wood chip bits to the agar I poured about a week ago and this is what is growing:






Should I let them grow some more or transfer a piece to some clean agar?


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InvisibleVirginiaGentleman
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Re: First time agar, azures MS syringe, which media should I use? [Re: Pestile]
    #18858053 - 09/18/13 01:25 PM (10 years, 4 months ago)

Looks clean, no major sectoring. If you want to transfer its ready, I mostly go by the moisture of the agar - if its peeling away from the sides its beginning to dry and should be transfered in a few days.

Maybe keep a culture in cold storage?


To clear things up, what's your goal for the agar work?


--------------------
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Edited by VirginiaGentleman (09/18/13 01:27 PM)


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InvisiblePestile

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Re: First time agar, azures MS syringe, which media should I use? [Re: VirginiaGentleman]
    #18858359 - 09/18/13 02:24 PM (10 years, 4 months ago)

I want to inoculate some rye with it and use that on some wood chips.
I want to make a couple beds when spring arrives, until then I need to buff up my substrate.
I plan on transferring these one more time to "clean" them, and then transfer once more, these I'll use for the rye.


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InvisibleVirginiaGentleman
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Re: First time agar, azures MS syringe, which media should I use? [Re: Pestile]
    #18860050 - 09/18/13 08:01 PM (10 years, 4 months ago)

Depending on your time frame, you can pull some grain size transfers from those cultures to produce more plates for inoc'ing rye. In the mean time the plates you pulled transfers from will continue growing and will be good for dropping in rye when you see fit.

Try going from grain to egg carton cuttings then to wood. The egg cartons are very permeable for the mycelium to quickly work through it and "adjust" its diet. My woodies have loved it so far and take to wood much quicker than my direct to wood efforts.


About keeping a cold culture, its an easy piece of mind if SHTF, that way you don't have to reclean from prints :mushroomgrow:


--------------------
Ps. Allenii, Arcana, Azure, Baeos, Caerulipes, Cyan, Ovoid, Subreg, etc FT - Only seeking Ps. woodlovers - legal USA + CAN - Intended for microscopy use only


Edited by VirginiaGentleman (09/18/13 08:05 PM)


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Offlinecoffeehead
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Re: First time agar, azures MS syringe, which media should I use? [Re: blojo02184]
    #18860399 - 09/18/13 09:18 PM (10 years, 4 months ago)

I've had bad luck with PDA but I think the dishes might have been too dry. Another top poster here recommends MEA. Potato based agar recipes are supposed to work well, also.

If it were me, I'd talk to the vendor about possibly getting a replacement. Or I'd just buy another. Is it really worth the aggravation and work? After all, it's only MS -- it's not like an isolate you want to perpetuate.

I have at least one MS of azurescens left, some ELME and agar. I plan on inoculating some of the short, fat jars with MEA as soon as I can get around to it. I'll post here if I succeed just because it seems like nobody knows much about azurescens -- or wood lovers in general.

If you know anything about fruiting and harvesting azurescens I'd greatly appreciate any advice. I posted a thread asking for help but got no replies.

Here's a link to my post.

Good luck.


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Re: First time agar, azures MS syringe, which media should I use? [Re: VirginiaGentleman]
    #18860499 - 09/18/13 09:35 PM (10 years, 4 months ago)

Quote:

VirginiaGentleman said:
Depending on your time frame, you can pull some grain size transfers from those cultures to produce more plates for inoc'ing rye. In the mean time the plates you pulled transfers from will continue growing and will be good for dropping in rye when you see fit.

Try going from grain to egg carton cuttings then to wood. The egg cartons are very permeable for the mycelium to quickly work through it and "adjust" its diet. My woodies have loved it so far and take to wood much quicker than my direct to wood efforts.


About keeping a cold culture, its an easy piece of mind if SHTF, that way you don't have to reclean from prints :mushroomgrow:




Thanks for the egg carton tip. I've had success adding from 20 - 40% (by volume) moist wood chips to the rye. For me, they transfer more quickly to the bulk chips than straight rye.


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Re: First time agar, azures MS syringe, which media should I use? [Re: coffeehead]
    #18860516 - 09/18/13 09:39 PM (10 years, 4 months ago)

Quote:

coffeehead said:


Thanks for the egg carton tip. I've had success adding from 20 - 40% (by volume) moist wood chips to the rye. For me, they transfer more quickly to the bulk chips than straight rye.





On my long sacred journeys deep into the shroomery search engine i find what i need!






Eventually..:gethigh:


But yeah, gradualism is essential for the jar style until you're ready for 1L+ of spawn


--------------------
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Edited by VirginiaGentleman (09/18/13 09:40 PM)


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