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Offlinetedoro
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Second transfer on soft agar... how does it look?
    #26418104 - 01/05/20 06:23 PM (4 years, 1 month ago)

I am starting afresh and started from spores ~ this is the second transfer. Golden Teacher. Malt agar made soft to assist blending LI.



a day later, here are a few more of my transfers, and they are starting to whiten up in the clear area...


Edited by tedoro (01/06/20 11:43 AM)


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Offlinezensane
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Re: Second transfer on agar... how does it look? [Re: tedoro]
    #26418219 - 01/05/20 07:28 PM (4 years, 1 month ago)

For me, when there is a distinct, liquified line in front of what looks like mycelium spreading, it is always pin mold or some type of bacterial contamination.  It is a bit early to tell if there is contamination on the first dish.  In 2 or 3 days' time, it will be more evident.

Take a look at this as an example:
Pin Mold

It is exactly what my dishes will do if they have pin mold.
I became quite the pin mold farmer thinking I was transferring mycelium.

If there is healthy mycelium, it will be leading the charge across the dish with nothing notable in front of it.


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Offlinemagicschoolbus
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Re: Second transfer on agar... how does it look? [Re: zensane]
    #26418251 - 01/05/20 07:45 PM (4 years, 1 month ago)

I would recommend taking more than 1 transfer at at time if you can. If you're short on plates, you could place them in the shape of a triangle, since you'll only be letting them grow to about the size of a nickel there will be plenty of room for 3 cultures to be transferred on to one plate.

Transferring several of the best pieces of tissue you can find will increase your chances of having at least one viable culture that can again be transferred. Also, doing only one transfer might limit your potential of hitting the genetic jackpot


--------------------
:trippinballs::trippinballs:


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Offlinetedoro
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Re: Second transfer on agar... how does it look? [Re: magicschoolbus]
    #26418276 - 01/05/20 07:57 PM (4 years, 1 month ago)

I did three separate spore starts... and I have many transfers of each. They all have this look though. This is just one petri dish.

I am almost positive this is not pin mold. My initial plate is weeks old now. I havent seen pin mold here in years.


--------------------
--------------------
Deep pour soft agar plates-->bags of WBS-->Low Profile Monos
Clean spawn thread | Put a thermometer on your PC


Edited by tedoro (01/05/20 07:58 PM)


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Offlineinsutama
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Re: Second transfer on agar... how does it look? [Re: tedoro]
    #26418287 - 01/05/20 08:02 PM (4 years, 1 month ago)

it seems my plate always have that clear growth in front of the mycelium also


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Offlinetedoro
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Re: Second transfer on agar... how does it look? [Re: tedoro]
    #26418288 - 01/05/20 08:03 PM (4 years, 1 month ago)

In a few days, this translucent outer ring will turn white, as I saw on the first transfer.

So this is not mycelia for sure?


--------------------
--------------------
Deep pour soft agar plates-->bags of WBS-->Low Profile Monos
Clean spawn thread | Put a thermometer on your PC


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Offlinetedoro
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Re: Second transfer on agar... how does it look? [Re: tedoro]
    #26418300 - 01/05/20 08:06 PM (4 years, 1 month ago)

And don't worry about my feelings here... I just need to know if I need to start fresh again with spores. It is strange that all three spore starts (from separate spore prints) look like this on their second transfer.

It should also be noted that I make softer agar as to make LI blending easier.


--------------------
--------------------
Deep pour soft agar plates-->bags of WBS-->Low Profile Monos
Clean spawn thread | Put a thermometer on your PC


Edited by tedoro (01/05/20 08:08 PM)


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Offlinetedoro
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Re: Second transfer on agar... how does it look? [Re: tedoro]
    #26418351 - 01/05/20 08:27 PM (4 years, 1 month ago)



Here is another picture of another transfer. They all look similar. These pictures are hard to shoot!


--------------------
--------------------
Deep pour soft agar plates-->bags of WBS-->Low Profile Monos
Clean spawn thread | Put a thermometer on your PC


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Offlinetedoro
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Re: Second transfer on agar... how does it look? [Re: insutama]
    #26418386 - 01/05/20 08:48 PM (4 years, 1 month ago)

Quote:

insutama said:
it seems my plate always have that clear growth in front of the mycelium also




Question: Do you get second and third flushes?


--------------------
--------------------
Deep pour soft agar plates-->bags of WBS-->Low Profile Monos
Clean spawn thread | Put a thermometer on your PC


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Offlinetedoro
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Re: Second transfer on agar... how does it look? [Re: tedoro]
    #26419178 - 01/06/20 11:47 AM (4 years, 1 month ago)

Its a day later and I just took another picture of the dish(3rd picture), and a few other dishes. We can see them whitening up. So my question is: Is this plate contaminated, or is this just a squirrelly new ms plate growing on soft agar? I question it being a contam... because it seems unlikely that all three of my spore print plates (from separate prints) would contam the same way.



--------------------
--------------------
Deep pour soft agar plates-->bags of WBS-->Low Profile Monos
Clean spawn thread | Put a thermometer on your PC


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Offlinezensane
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Re: Second transfer on agar... how does it look? [Re: tedoro]
    #26419865 - 01/06/20 07:20 PM (4 years, 1 month ago)

Here are a few pics that may help describe what I was trying to communicate.
Good mycelium has a very distinct boundary.
These plates are about 12 days further matured than where your plates are.
It doesn't look watery or disturbed past the leading edge.

I hope this helps.





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OfflineRandyFeller
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Re: Second transfer on agar... how does it look? [Re: zensane]
    #26419929 - 01/06/20 07:50 PM (4 years, 1 month ago)

Mycelium seems to grow differently on the soft agar. Not sure if it's because of the super low nute ratio or if the mycelium is growing thru the soft agar beneath the surface.

I've had this same question running thru my head here lately playing with the soft agar..so much that I am now running normal recipe agar for plates I won't be putting to grain so that I can be positive I'm selecting good myc. I now transfer the "normal" looking mycelium to soft agar for A2g transfers


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Offlineinsutama
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Re: Second transfer on agar... how does it look? [Re: RandyFeller]
    #26419953 - 01/06/20 08:09 PM (4 years, 1 month ago)

My agar is also soft so I'm thinking the soft agar is maybe the reason for the translucent outer ring on my plates


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Invisiblerickyswamps
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Re: Second transfer on agar... how does it look? [Re: insutama]
    #26420022 - 01/06/20 08:53 PM (4 years, 1 month ago)

Its too early to tell what is going on.  Give them some time to grow and then post updates.


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OnlineA.k.aM
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Re: Second transfer on agar... how does it look? [Re: rickyswamps]
    #26420173 - 01/06/20 10:44 PM (4 years, 1 month ago)

There was a conversation about this somewhere fairly recently, the general consensus was that the clear circle just happens sometimes.

They all look fine to me. If it was mold you would know by now and I haven’t seen bacteria grow that uniform or without bubbling up or being wet and runny.

Prob just genetic/agar combo.


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


Edited by A.k.a (01/06/20 10:44 PM)


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Offlinetedoro
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Re: Second transfer on agar... how does it look? [Re: A.k.a]
    #26420992 - 01/07/20 12:41 PM (4 years, 1 month ago)


I really appreciate you all chiming in.

These are the three versions of my Golden Teacher I am cleaning up shot just now...


I am feeling they are clean and just sorta diving into the agar a bit at the beginning.

ZenSane, I agree with what you are pointing out, but perhaps soft agar can cause this ring. They will look fabulous here in a day or two. OOOOrrrr.... its a contam that the mycelia is taking over....


--------------------
--------------------
Deep pour soft agar plates-->bags of WBS-->Low Profile Monos
Clean spawn thread | Put a thermometer on your PC


Edited by tedoro (01/07/20 01:11 PM)


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InvisibleSpunkyMonkey88
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Re: Second transfer on agar... how does it look? [Re: tedoro]
    #26421056 - 01/07/20 01:05 PM (4 years, 1 month ago)

I really think you are ok. You should just give them a couple of days to see how they play out but my guess is they are ok...

When I have a contaminate that makes a translucent ring infront of the mushroom mycelium, it is never uniform to the transfer wedge and does it's own thing...


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Offlinetedoro
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Re: Second transfer on agar... how does it look? [Re: SpunkyMonkey88]
    #26421074 - 01/07/20 01:10 PM (4 years, 1 month ago)

I like hearing its likely good, but if there is such a thing as a contam that subtly hitches a ride on mycelia, then I'm still unsettled by what I see.

RandyFeller: I do like the idea of making normal and soft agar to remove that variable. Have you found that the clear ring disappears on the normal agar?


--------------------
--------------------
Deep pour soft agar plates-->bags of WBS-->Low Profile Monos
Clean spawn thread | Put a thermometer on your PC


Edited by tedoro (01/07/20 01:11 PM)


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InvisibleAyePlusS
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Re: Second transfer on agar... how does it look? [Re: tedoro]
    #26421092 - 01/07/20 01:20 PM (4 years, 1 month ago)

Try a transfer to water agar or a hot pour to be sure there isnt anything hitching along, and there definitely are contams that hide and intertwine with myc. Pin molds and bacteria can both be pretty tricky to spot, and oh ya ted check your PM’s. I can send you anAA+ or KSSS print if you want thats all i have extra rn.


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