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Offlinespore-ty
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Some dishes
    #23707280 - 10/04/16 03:41 PM (7 years, 3 months ago)

Hello everyone so i have 3 b+ plates one of a actual pin,
One of just mycelium from a substrate with knots on it,
And another of a pin

These were my first plates ever i pced my pyrex bottle for 45 mins waited to cool then poured in sab waited to cool and inoculated

got my movements down for the plates i did two days ago started from the bottom in stacks of 3 these plates much more smoother will post pics of those when there are updates

But ya these are pics today before transfer




Edited by spore-ty (10/04/16 03:46 PM)


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InvisiblemushboyMDiscord
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Re: Some dishes [Re: spore-ty] * 1
    #23707289 - 10/04/16 03:44 PM (7 years, 3 months ago)

looks decent. trim that nail


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Offlinespore-ty
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Re: Some dishes [Re: mushboy]
    #23707302 - 10/04/16 03:49 PM (7 years, 3 months ago)

When i do agar to agar its fine to cut into the actual agar right?

And
Quote:

mushboy said:
trim that nail




yes massa


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Offlinethebug76
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Re: Some dishes [Re: spore-ty]
    #23707329 - 10/04/16 04:00 PM (7 years, 3 months ago)

Yes, its much easier than trying to get tthe mycelium off the top of it. Cut the agar into wedges and use those to transfer.


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OfflineTheLongComma
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Re: Some dishes [Re: spore-ty]
    #23707332 - 10/04/16 04:03 PM (7 years, 3 months ago)

Quote:

spore-ty said:
When i do agar to agar its fine to cut into the actual agar right?





Yes, you cut the chunk and stab it.

Your 4th picture looks like it has bacteria surrounding the myc. All of that opaque blotch is what I'm talking about


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Offlinespore-ty
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Re: Some dishes [Re: TheLongComma]
    #23707365 - 10/04/16 04:15 PM (7 years, 3 months ago)

They all have some contams on them i planned on it


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OfflinePeacefrog
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Re: Some dishes [Re: spore-ty]
    #23707440 - 10/04/16 04:48 PM (7 years, 3 months ago)

Cutting very tiny pieces of mycelium to transfer from plate to plate is the best practice if you are either cleaning up or isolating. And I would not stab the wedge when transferring to a new agar plate. Very quickly, carefully cut the wedge, and just barely position it on the edge of your scalpel. Then upon transferring to a new dish, you will only open the receiving dish for a second or two and just touch the tip of the scalpel to the agar and roll slightly. The wedge should come right off in a very quick and deliberate motion.


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Offlinespore-ty
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Re: Some dishes [Re: Peacefrog]
    #23707476 - 10/04/16 05:03 PM (7 years, 3 months ago)

Quote:

Peacefrog said:
Cutting very tiny pieces of mycelium to transfer from plate to plate is the best practice if you are either cleaning up or isolating. And I would not stab the wedge when transferring to a new agar plate. Very quickly, carefully cut the wedge, and just barely position it on the edge of your scalpel. Then upon transferring to a new dish, you will only open the receiving dish for a second or two and just touch the tip of the scalpel to the agar and roll slightly. The wedge should come right off in a very quick and deliberate motion.





I knew about the small wedges but every thing else no clue about thanks a ton

Question how long does it take the agar wedge to begin to recolonize?


Edited by spore-ty (10/04/16 05:03 PM)


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OfflinePeacefrog
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Re: Some dishes [Re: spore-ty]
    #23707554 - 10/04/16 05:25 PM (7 years, 3 months ago)

For cubes, you should notice the wedge starting to fuzz up in 24-36 hours and usually will "dig in to" the new agar within 2-4 days on average.


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OfflineGreg
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Re: Some dishes [Re: Peacefrog]
    #23707643 - 10/04/16 05:56 PM (7 years, 3 months ago)

I see a film of bacteria on a couple of those plates.

I outlined it in this pic, the original is next to it for comparison:


Am I going crazy or do other people see it too?


Edited by Greg (10/04/16 06:08 PM)


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InvisibleComebackKid
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Re: Some dishes [Re: Peacefrog]
    #23707652 - 10/04/16 06:05 PM (7 years, 3 months ago)

Quote:

Peacefrog said:
Cutting very tiny pieces of mycelium to transfer from plate to plate is the best practice if you are either cleaning up or isolating. And I would not stab the wedge when transferring to a new agar plate. Very quickly, carefully cut the wedge, and just barely position it on the edge of your scalpel. Then upon transferring to a new dish, you will only open the receiving dish for a second or two and just touch the tip of the scalpel to the agar and roll slightly. The wedge should come right off in a very quick and deliberate motion.



THANKYOU!!
So obvious probably but I was having so much trouble getting the scuwered wedge off my scalpel that I began to just wipe the blade on the edge of the plate and position to the center later.
Haven't noticed more contams with this method yet but every time I did that it just felt so wrong and felt like it would eventually catch up with me somehow.


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:amanita2: Substrate surface conditions / Monotub prep and care :sporedrop:

Look around you... Everything you see exists inside the mind.
Consciousness, the awareness that is experiencing this mind,
is peering in from outside the universe.

Our individual experiences are all part of the universe's experience of itself


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OfflineKenetic
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Re: Some dishes [Re: ComebackKid]
    #23708068 - 10/04/16 08:38 PM (7 years, 3 months ago)

Don't touch the side of the dish with your scalpel, this is just begging for contams. 

I like to position the wedge I'm transferring onto the new dish, tilt the scalpel at a slight angle to the side, and move my scalpel sideways while applying slight downward pressure.  This helps remove the wedge fairly easily.  Also, don't stab the wedge too hard when you're picking it up. 

And that's a shit-ton of bacteria on the one plate, how did that happen?  I've only seen bacteria colonies converge like that if they were allowed to grow out too long.


--------------------
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OfflineGreg
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Re: Some dishes [Re: spore-ty]
    #23708075 - 10/04/16 08:39 PM (7 years, 3 months ago)

Also, one of the other plates has bacteria all around the rim.
This one:


Edit: and maybe some mold near the clone too.


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OfflineKenetic
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Re: Some dishes [Re: Kenetic]
    #23708081 - 10/04/16 08:41 PM (7 years, 3 months ago)

Also, you don't need to pc agar for 45 minutes.  Try 20-25 at 15 psi


--------------------
Todo Cambia
   

               
                                                :cookiemonster::elmo:



DMT said: Everyone know's me, they just don't know it yet


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Offlinespore-ty
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Re: Some dishes [Re: Greg]
    #23708377 - 10/04/16 10:02 PM (7 years, 3 months ago)

Haha no your not going crazy your right
Quote:

Greg said:
I see a film of bacteria on a couple of those plates.

I outlined it in this pic, the original is next to it for comparison:


Am I going crazy or do other people see it too?





But even with all these contams are these still viable for tranfers?


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Offlinespore-ty
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Re: Some dishes [Re: Greg]
    #23708472 - 10/04/16 10:39 PM (7 years, 3 months ago)

Quote:

Greg said:
Also, one of the other plates has bacteria all around the rim.
This one:


Edit: and maybe some mold near the clone too.





Correct again! Lol your good


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Offlinespore-ty
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Re: Some dishes [Re: Kenetic]
    #23708477 - 10/04/16 10:42 PM (7 years, 3 months ago)

Quote:

kenetic said:
Don't touch the side of the dish with your scalpel, this is just begging for contams. 

I like to position the wedge I'm transferring onto the new dish, tilt the scalpel at a slight angle to the side, and move my scalpel sideways while applying slight downward pressure.  This helps remove the wedge fairly easily.  Also, don't stab the wedge too hard when you're picking it up. 

And that's a shit-ton of bacteria on the one plate, how did that happen?  I've only seen bacteria colonies converge like that if they were allowed to grow out too long.




It was my first time pouring dishes and using a sab lol..


Sorry for the triple posts pretty stoned and just kept losing train of thought wont happen again


Edited by spore-ty (10/04/16 10:43 PM)


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OfflinePeacefrog
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Re: Some dishes [Re: spore-ty]
    #23708769 - 10/05/16 01:48 AM (7 years, 3 months ago)

Yes you should be able to clean those up. Look up sandwich agar tek. I have never personally tried it, but it sounds like a very good means of cleaning up a bacterial dish.

I just prefer to clean it up the old fashioned way. One trick you can use, if going this route, is to use the back edge of your scalpel or an inoculating loop to gently rub some healthy mycelium as far away from the contaminate as possible and transfer to a new dish. Do this to several plates and continue to clean up if necessary.

Good luck and nice job for your first trials with agar.


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Offlinespore-ty
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Re: Some dishes [Re: Peacefrog]
    #23708964 - 10/05/16 05:55 AM (7 years, 3 months ago)

Quote:

Peacefrog said:
Yes you should be able to clean those up. Look up sandwich agar tek. I have never personally tried it, but it sounds like a very good means of cleaning up a bacterial dish.

I just prefer to clean it up the old fashioned way. One trick you can use, if going this route, is to use the back edge of your scalpel or an inoculating loop to gently rub some healthy mycelium as far away from the contaminate as possible and transfer to a new dish. Do this to several plates and continue to clean up if necessary.

Good luck and nice job for your first trials with agar.





Haha thanks mann
And ill remember that later today when im doing some transfers


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