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OfflineDr. Sporesly
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Registered: 05/20/20
Posts: 72
Last seen: 1 month, 18 days
Which agar plates / sectors to move to grain?
    #26836036 - 07/21/20 12:41 PM (3 years, 6 months ago)

Hey all,

After a bunch of transfers and isolations over the past month or two I'm finally ready to move to grains (12 jars total).

From the pictures below, would anyone be able to suggest which plate(s)/sectors I should use? Please note plate 6 has some bacterial growth on the side.

I would like to keep one or two plates going to continue isolating the mycelium, so if you can suggest which one to keep that would be great as well.

Also, do my grains need to get to room temperature after PC or how long should they sit before before I throw the agar into them?

Thanks!



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Invisiblealaskappalachian
Entitiologist


Registered: 10/22/19
Posts: 1,674
Loc: The 49th Dimension
Re: Which agar plates / sectors to move to grain? [Re: Dr. Sporesly]
    #26836057 - 07/21/20 12:56 PM (3 years, 6 months ago)

Plate  1 I'd grab the better organized growth @ 7:00
Plate 2 grab from the leading edge of the rhizo sector
Plate 3 same  (12:00)
Plate 4 grab from the rhizo but a bit in from the edge of the plate
Plate 5 I'd go with 7:00
Plate 6 I'd take from the 5:00 area actually.
Most of those plates have some issues with bacteria btw. Keep transferring until you get some uniformity in growth.
If I was going to use one of those I'd use plate 1.  You can use grain jars like 6hrs after PC IME. 


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"First we build the tools, then they build us."

THE 49th MYCOJOURNAL: Exotics, Auroras, and Entities


Edited by alaskappalachian (07/21/20 01:07 PM)


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OfflineDr. Sporesly
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Registered: 05/20/20
Posts: 72
Last seen: 1 month, 18 days
Re: Which agar plates / sectors to move to grain? [Re: alaskappalachian]
    #26836108 - 07/21/20 01:30 PM (3 years, 6 months ago)

Quote:

alaskappalachian said:
Plate  1 I'd grab the better organized growth @ 7:00
Plate 2 grab from the leading edge of the rhizo sector
Plate 3 same  (12:00)
Plate 4 grab from the rhizo but a bit in from the edge of the plate
Plate 5 I'd go with 7:00
Plate 6 I'd take from the 5:00 area actually.
Most of those plates have some issues with bacteria btw. Keep transferring until you get some uniformity in growth.
If I was going to use one of those I'd use plate 1.  You can use grain jars like 6hrs after PC IME. 





Thanks I appreciate it!

1. Would you recommend I ignore inoculating the grain now and just do more transfers? I don't see any bacteria on the other plates, can you call out where you see it, or what is the sign for it?

2. For cultivation based on agar, should I be looking for Rhizo or Tomentose? I've read so much conflicting information so I'm not really sure what to isolate, I've just been isolating what looks to be the fastest growing part of the agar.

3. Would it be best to just start with one jar and doing G2G? I've already Pc'd all my jars so... to keep them sterile can i just store them until they're ready, or should i just re-PC when I'm ready to use

Thanks!!!


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OfflineTheGreatGig
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Registered: 07/02/20
Posts: 45
Last seen: 3 years, 4 months
Re: Which agar plates / sectors to move to grain? [Re: alaskappalachian]
    #26836129 - 07/21/20 01:40 PM (3 years, 6 months ago)

Quote:

alaskappalachian said:
Most of those plates have some issues with bacteria btw. 




Just out of curiosity where do you see bacteria on most plates? Plate 6 is obvious but I cant see any wrong in the other plates. From what I see they are just trapped air bubbles. Agar noob here as well


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OfflineDr. Sporesly
Papa Smurf

Registered: 05/20/20
Posts: 72
Last seen: 1 month, 18 days
Re: Which agar plates / sectors to move to grain? [Re: TheGreatGig]
    #26836145 - 07/21/20 01:46 PM (3 years, 6 months ago)

Quote:

TheGreatGig said:
Quote:

alaskappalachian said:
Most of those plates have some issues with bacteria btw. 




Just out of curiosity where do you see bacteria on most plates? Plate 6 is obvious but I cant see any wrong in the other plates. From what I see they are just trapped air bubbles. Agar noob here as well




That's my question too .. I don't see any and I have other plates that are sitting now for over a month (kept them because I want to see if they pin and can clone) and they have zero bacteria either.

Don't get me wrong, these plates are sloppy because of the air bubbles .. tried something new and kinda shook/moved them in a circle and it fucked them bad lol


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OfflineTheGreatGig
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Registered: 07/02/20
Posts: 45
Last seen: 3 years, 4 months
Re: Which agar plates / sectors to move to grain? [Re: Dr. Sporesly]
    #26836148 - 07/21/20 01:48 PM (3 years, 6 months ago)

Dont re-PC jars just leave them in the pc until use or store them in a cool and clean place. As long as you dont open them they should remain sterile, but use them sooner rather than later.

I would nocc up a few jars at least to test out your sterile technique and mycelium


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Invisiblealaskappalachian
Entitiologist


Registered: 10/22/19
Posts: 1,674
Loc: The 49th Dimension
Re: Which agar plates / sectors to move to grain? [Re: TheGreatGig]
    #26836160 - 07/21/20 01:54 PM (3 years, 6 months ago)

Sometimes bacterial interactions can be seen in the way the mycelium grows (such as radically different sectors erupting from disorganized growth, or disorganized growth).  Basically you're shooting for good uniformity all around, and rhizo growth is preferable (although I've gone full-send on many tomentose plates).  What really matters is well organized, uniform growth.  Your plates don't look bad, but I'd definitely transfer them once or twice more.  A week won't hurt those grain jars.  Like I said: maybe drop the first plate if you want to make something happen and if it's good, g2g that bugger.  That's what I'd do actually.  You could drop most of these and get some growth, no prob; but you'll see a huge benefit from another transfer or two.  I spend most of my time with agar, and let g2g make up  for lost time.  You're doing great.  Go ahead and drop that more uniform plate 1 though.  Also store unused jars on shelf.  Not in the PC.


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


"First we build the tools, then they build us."

THE 49th MYCOJOURNAL: Exotics, Auroras, and Entities


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OfflineDr. Sporesly
Papa Smurf

Registered: 05/20/20
Posts: 72
Last seen: 1 month, 18 days
Re: Which agar plates / sectors to move to grain? [Re: alaskappalachian]
    #26836218 - 07/21/20 02:25 PM (3 years, 6 months ago)

Quote:

alaskappalachian said:
Sometimes bacterial interactions can be seen in the way the mycelium grows (such as radically different sectors erupting from disorganized growth, or disorganized growth).  Basically you're shooting for good uniformity all around, and rhizo growth is preferable (although I've gone full-send on many tomentose plates).  What really matters is well organized, uniform growth.  Your plates don't look bad, but I'd definitely transfer them once or twice more.  A week won't hurt those grain jars.  Like I said: maybe drop the first plate if you want to make something happen and if it's good, g2g that bugger.  That's what I'd do actually.  You could drop most of these and get some growth, no prob; but you'll see a huge benefit from another transfer or two.  I spend most of my time with agar, and let g2g make up  for lost time.  You're doing great.  Go ahead and drop that more uniform plate 1 though.  Also store unused jars on shelf.  Not in the PC.




Appreciate it – thanks!

Would I be able to cut plate 1 in half and use it across 2 jars?

Once I open the PC I'll have 5 jars left over. Should I just store them on a shelf and PC again when I need them in a week, or just get more grain and do it again?

Also, here are 3 plates that are maybe a month old now – decided to keep a few based on a Bod post where he said you can pin a plate and clone a pin.

R3 and R4 are Rhizo and T1 is Tementose, right?

I guess I can actually drop any of these in a jar as well if you think they're decent enough?



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Invisiblealaskappalachian
Entitiologist


Registered: 10/22/19
Posts: 1,674
Loc: The 49th Dimension
Re: Which agar plates / sectors to move to grain? [Re: Dr. Sporesly]
    #26836300 - 07/21/20 03:12 PM (3 years, 6 months ago)

Yep.  You can do one plate to two jars. Slower, but just fine.  All those show rhizomorphic growth btw.  R3 and R4 are just more ropey.  You could drop a wedge from R4 if you want to experiment, staying away from the edge and omitting the bit of growth between 6-7 o'clock.  Or just hold out for pins.  I sometimes drop a wedge off older plates when I'm bored.  If they have been growing all that time and don't have mold issues, you can see what happens.  It's a bit more wise to wait for a plate pin though.  I'm sure you'll get one or two if you wait.  Realistically, you can drop a wedge off any of those and get some kind of growth and select your best jars for g2g.  If you have grain and time, go for it; but working retroactively and pulling from those old plates just to tinker, I'd pull from R4.  Just set your excess jars on the shelf.  If your filter is good you'll be fine for a week.  Between g2g and shaking, that amount of time won't matter with clean myc.  You got this, bud.


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


"First we build the tools, then they build us."

THE 49th MYCOJOURNAL: Exotics, Auroras, and Entities


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