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Offlineyojay50
always learning


Registered: 04/21/17
Posts: 342
Last seen: 2 months, 20 days
questions regarding spores and tissue on agar
    #26453790 - 01/26/20 03:41 PM (4 years, 22 days ago)

when i first started this i made 8 pasty plates and 1 small jar of grain.

i inoculated everything at around the same time, now its like 2 weeks later and the grain jar has some growth but the plates not really except for one which contamed out.

i eyeballed the agar amount and i know for sure it was too hard when it was done in the pc.  i made 4 new plates about 4 days ago, they were WAY softer than the first batch. but not rolling around goopy soft, it was firm but also gave way ez.

i put oyster mushrooms tissue on 2 of the new plates and also put spores on one. one plate is still uninoculated.

the oyster tissue i put on the plates did the thing where it "melts" and then myc starts growing out from it.

was wondering if that means i can just start a grain jar with tissue from a mushroom.  ones you can get that are fresh. who knows.

whats a good agar recipe for starting spores.


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OfflineMorgan823
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Registered: 01/23/20
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Last seen: 4 years, 22 days
Re: questions regarding spores and tissue on agar [Re: yojay50]
    #26453966 - 01/26/20 05:56 PM (4 years, 22 days ago)

If you’re doing work with agar plates, it’s necessary to have a flow good to work in front of and be knowledgeable in sterile technique.  I often times use a simple potato broth recipe of 20 grams agar, to 1 liter of liquid (potato broth (the water from boiling potatoes) and added water if there is not enough broth).  Sometimes I add nutritional yeast and peptones.  As far as tissue and spore culturing onto plates goes, I recommend reading up on proper technique.  Both Tradd Cotter’s book “Mushroom Farming and Mycoremediation” and Paul Stamets “Growing Gourmet and Medicinal Mushrooms” have good sections on this.  No, you cannot tissue culture directly into a jar.


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OfflineA.k.aM
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Registered: 10/27/19
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Re: questions regarding spores and tissue on agar [Re: Morgan823]
    #26453976 - 01/26/20 06:02 PM (4 years, 22 days ago)

You don’t need a hood, just a still air box.

And you definitely can drop tissue into a jar if it’s clean.


--------------------
LAGM2020


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Offlineyojay50
always learning


Registered: 04/21/17
Posts: 342
Last seen: 2 months, 20 days
Re: questions regarding spores and tissue on agar [Re: Morgan823]
    #26454779 - 01/27/20 08:53 AM (4 years, 21 days ago)

Quote:

Morgan823 said:
If you’re doing work with agar plates, it’s necessary to have a flow good to work in front of and be knowledgeable in sterile technique.  I often times use a simple potato broth recipe of 20 grams agar, to 1 liter of liquid (potato broth (the water from boiling potatoes) and added water if there is not enough broth).  Sometimes I add nutritional yeast and peptones.  As far as tissue and spore culturing onto plates goes, I recommend reading up on proper technique.  Both Tradd Cotter’s book “Mushroom Farming and Mycoremediation” and Paul Stamets “Growing Gourmet and Medicinal Mushrooms” have good sections on this.  No, you cannot tissue culture directly into a jar.




Your first post was that comment? Hmmmm I smell fishhhhhh. I have experience with sterile technique. Only one of my plates hard contaminated.


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Offlineyojay50
always learning


Registered: 04/21/17
Posts: 342
Last seen: 2 months, 20 days
Re: questions regarding spores and tissue on agar [Re: A.k.a]
    #26454782 - 01/27/20 08:55 AM (4 years, 21 days ago)

Quote:

A.k.a said:
You don’t need a hood, just a still air box.

And you definitely can drop tissue into a jar if it’s clean.




That's what I'm thinking, if the tissue sample is clean (from inside the mushroom)

I can just drop that bad boy on grain like we drop it on agar!

Also my plates were too dry but I think I see a bit of myc. My grain jar has growth tho and it's nice white and mycy


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OfflinecronicrFacebook
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Re: questions regarding spores and tissue on agar [Re: yojay50]
    #26454812 - 01/27/20 09:16 AM (4 years, 21 days ago)

U could but far safer to go to agar first, u already have it you may as well use it. Any standard recipe is fine for using spores, softer agar may be more forgiving but not a deal breaker especially with no pours


--------------------

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Offlineyojay50
always learning


Registered: 04/21/17
Posts: 342
Last seen: 2 months, 20 days
Re: questions regarding spores and tissue on agar [Re: cronicr]
    #26455046 - 01/27/20 12:02 PM (4 years, 21 days ago)

Quote:

cronicr said:
U could but far safer to go to agar first, u already have it you may as well use it. Any standard recipe is fine for using spores, softer agar may be more forgiving but not a deal breaker especially with no pours



Quote:

cronicr said:
U could but far safer to go to agar first, u already have it you may as well use it. Any standard recipe is fine for using spores, softer agar may be more forgiving but not a deal breaker especially with no pours





do you know any standard recipes?

just weights and amounts.

i do sweet potato broth with pure unprocessed sugar cane sugar.

just need the amount of agar (grams) and everything else in grams


seeing my oyster mycelium spread makes tingle inside


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OfflineNorthman
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Registered: 01/27/20
Posts: 61
Last seen: 3 years, 1 month
Re: questions regarding spores and tissue on agar [Re: yojay50]
    #26455360 - 01/27/20 02:46 PM (4 years, 21 days ago)

400ml of water, 6 grams of light malt extract, 7 grams of agar. And I agree, SAB is fine for agar work. Flowhoods are great but not everyone wants to go that far. If using jars for agar, use half pints and turn the lid upside down before screwing the metal ring on before PCing to avoid the vacuum effect pulling contaminated air in when opening the jar.


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InvisibleGoombas69
Amateur Mycologist

Registered: 12/15/19
Posts: 15
Re: questions regarding spores and tissue on agar [Re: Northman]
    #26455680 - 01/27/20 05:54 PM (4 years, 21 days ago)

Quote:

Northman said:
400ml of water, 6 grams of light malt extract, 7 grams of agar. And I agree, SAB is fine for agar work. Flowhoods are great but not everyone wants to go that far. If using jars for agar, use half pints and turn the lid upside down before screwing the metal ring on before PCing to avoid the vacuum effect pulling contaminated air in when opening the jar.




Do you get much condensation with this recipe, and if so, how do you get rid of it? The most common recipe I have seen is 500ml water, 10g agar, 10g LME, but it seemed too dry/hard compared to premade petri dishes. (Your ratio would be 8.75g agar for 500ml water.) I'm tempted to try your recipe.


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OfflineNorthman
Stranger
Registered: 01/27/20
Posts: 61
Last seen: 3 years, 1 month
Re: questions regarding spores and tissue on agar [Re: Goombas69]
    #26455787 - 01/27/20 06:55 PM (4 years, 20 days ago)

When using petri dishes I never pour until it's under 119 degrees (just works for me). When using jars I follow this procedure and rarely have any condensation at all:

Mix agar in media bottle.

Microwave, shaking intermittently.

Pour into half pints jars. Turn lids upside down and screw metal ring on loosely. Put foil over top.

PC for 20 minutes at 15psi.

Quick vent (I've never had a problem with jars breaking) and remove the jars onto clean countertop.

Leave foil on loosely fitted lids until agar cools almost completely.

Tighten lids and remove foil.

Now if you get to the last step and notice some condensation, simple turn the jar upside down with the foil still over the lid and gently rock the jar side to side so that the water exits through the crack in the lid into the foil. Then tighten the lid with the jar still upside down. I've never had any contam issues doing it this way and I've been culturing oysters on agar using this method for well over a decade.


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InvisibleGoombas69
Amateur Mycologist

Registered: 12/15/19
Posts: 15
Re: questions regarding spores and tissue on agar [Re: Northman]
    #26455947 - 01/27/20 08:14 PM (4 years, 20 days ago)

Thank you so much Northman, I'm going to use your method. I really appreciate your response.


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