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Offlineboonquack
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Registered: 12/10/17
Posts: 11
Last seen: 4 years, 2 months
Agar - Wood Species Grow Plan, Starting from agar plate.
    #24912151 - 01/13/18 10:15 PM (7 years, 4 days ago)

New Project : Wood Species Culture
Starting Materials : Colonized Agar Plate
Lab Specs: 66 qt SAB, Presto 6qt PC
Goals :
1: Preserve Original Petri Culture (P1) as much as possible
2: Grow Out Mycelium to fruit.

Initial Plan:
1. Petri(P1)->Petri(P2) transfer of original plate: 4 plates done for statistical reduction of failure.
2. Reseal + Preserve Original Petri(P1) in refrigeration to preserve clean culture.

Wood Species Grow Plan (Low Budget)
1. Observe 4 plates(P2), select for growth/cleanliness.
2. Petris(P2) -> Bulk Innoculant ( P2->LC vs P2 to P3,4,5)
Not sure whether the best idea is to
A)attempt to make LC out of colonized petris (or if it would work for this lignicolous species)
B)keep growing out multiple agar plates.
C)agar to grain, then grain to grain transfer
Input would be helpful!
3. Bulk Innoculant -> Grain Spawn (Rye berries)
4. Grain Spawn -> Cardboard
5. Cardboard -> Outdoor containers w/ alder chips, pine/leaf litter, spawn and substrate casing. I have noticed that the outside seems to naturally grown mushrooms well, so I am going to attempt to use that to my advantage.

Pic Of Starting Sample (P1)

Where should i cut for the Petri transfers? Any tips for a first timer? I have an xacto and sharpened dab tools as surgical instruments.

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OfflinehamloafM
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Re: Agar - Wood Species Grow Plan, Starting from agar plate. [Re: boonquack]
    #24912162 - 01/13/18 10:23 PM (7 years, 4 days ago)

Agar to grain.
Grain to grain.
Grain to sterilzed woodchips.
Colonized woodchips to outdoor beds.


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Offlineboonquack
Institutionalized Education

Registered: 12/10/17
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Last seen: 4 years, 2 months
Re: Agar - Wood Species Grow Plan, Starting from agar plate. [Re: hamloaf]
    #24912205 - 01/13/18 10:50 PM (7 years, 4 days ago)

thank you for your quick reply. what size agar chunks for agar to grain?
I'm a complete noob with sterile technique. I understand going agar straight to grain for purposes of growing it out.  I suppose its possible to go grain to agar as well.  Are you talking chop the whole thing up and dump it in a quart jar, or just a piece of the pie.  would i be better off trying to inoculate a pint jar as a grain master, or a quart jar? 

my only qualm is that preserving the culture is important to me.  I will eventually make master slants.  i have some glass test tubes and wooden dowels.  thinking about getting purpose built culture tubes off amazon for ease, since i only have rubber stoppers for lids.  i want to reduce the chance of contaminating the initial sample (P1).

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OfflinehamloafM
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Re: Agar - Wood Species Grow Plan, Starting from agar plate. [Re: boonquack]
    #24912217 - 01/13/18 10:58 PM (7 years, 4 days ago)

Cut the culture into eighths.
It's possible.
Inoculate each jar using 1-2 wedges of agar.
Quart jars.


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How I Get Stuff done. - My Reference Guide. - My Grows.

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Offlineboonquack
Institutionalized Education

Registered: 12/10/17
Posts: 11
Last seen: 4 years, 2 months
Re: Agar - Wood Species Grow Plan, Starting from agar plate. [Re: hamloaf]
    #24912237 - 01/13/18 11:11 PM (7 years, 4 days ago)

Is gypsum recommended for the rye soak?
8/th pieces sound good. is it advisable to only use half the original culture in order to save it? or does the likelihood of contamination during the process make it a better idea to use all of the culture 1st time?  I was thinking of slicing 4 8th's off to do 4 jars, then saving the other half of the original culture?

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OfflinehamloafM
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Re: Agar - Wood Species Grow Plan, Starting from agar plate. [Re: boonquack]
    #24912261 - 01/13/18 11:23 PM (7 years, 4 days ago)

Not recommend, but doesn't hurt. 
Gypsum adds calcium and sulfate, as well as, prevents grains from clumping.
To preserve a culture transfer it on agar several times to ensure cleanliness, then slant it.


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How I Get Stuff done. - My Reference Guide. - My Grows.

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Offlineboonquack
Institutionalized Education

Registered: 12/10/17
Posts: 11
Last seen: 4 years, 2 months
Re: Agar - Wood Species Grow Plan, Starting from agar plate. [Re: boonquack]
    #24912262 - 01/13/18 11:23 PM (7 years, 4 days ago)

also, what about preparing an alder jar to use for colonization from agar vs grain?

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OfflinehamloafM
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Re: Agar - Wood Species Grow Plan, Starting from agar plate. [Re: boonquack]
    #24912272 - 01/13/18 11:26 PM (7 years, 4 days ago)

Grain provides the majority of nutrition, and will colonize the wood faster, and more evenly.


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How I Get Stuff done. - My Reference Guide. - My Grows.

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Offlineboonquack
Institutionalized Education

Registered: 12/10/17
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Last seen: 4 years, 2 months
Re: Agar - Wood Species Grow Plan, Starting from agar plate. [Re: hamloaf]
    #24912336 - 01/13/18 11:50 PM (7 years, 4 days ago)

So here is what I am thinking now.
P1->8th slices;
4 petris (P2)
4 rye grain (G1) jars (RR tek)

I want to do pint jars for grain due to my small PC, and to keep it small scale (for now) any reason not to, or precautionary tips?

for the petris, i will prepour and PC 2 glass petris(to ensure 2 reasonably sterile ones), and use 2 that i have ready (and hopefully arent contaminated, but it was a first try) for 4 total.

Hopefully 1 of the 8 slices will grow to be something beautiful.  Will a mixed grain/wood jar work?

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Offlinemushpunx
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Registered: 04/20/14
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Re: Agar - Wood Species Grow Plan, Starting from agar plate. [Re: boonquack]
    #24912852 - 01/14/18 07:38 AM (7 years, 3 days ago)

What species are you going to grow?


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Offlineboonquack
Institutionalized Education

Registered: 12/10/17
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Last seen: 4 years, 2 months
Re: Agar - Wood Species Grow Plan, Starting from agar plate. [Re: mushpunx]
    #24913454 - 01/14/18 12:31 PM (7 years, 3 days ago)

Not trying to be a jerk, just protecting "us."  We'd rather not discuss specifics,  but it is  a saprotrophic, wood dwelling species. Growth parameters are known and it has been cultured invitro and in outdoor beds. My question primarily relates to my plan and my limited experience with sterile technique, and maximizing my chances of success.  I don't want to fuck it up.  Am I just being a little bitch about slicing some sea jello?

TL:DR What would you do given a plate, with limited facility and experience in order to maximize chances of grow success and hopefully preserve the culture for future use?

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Offlinemushpunx
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Re: Agar - Wood Species Grow Plan, Starting from agar plate. [Re: boonquack]
    #24913485 - 01/14/18 12:50 PM (7 years, 3 days ago)

Nothing wrong with pints, just means more jars is all.

If you want to keep the original plate as your master, take a couple transfers to new plates to grow out.
Then wrap the edge + place in a ziplock, store in the fridge.

That way you have a master and a second working plate in cold storage and a working plate grown out to inoculate with


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