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the dagda
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Registered: 08/06/21
Posts: 30
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almost time for my first agar transfer i think. any advice?
#27460689 - 09/08/21 04:39 PM (3 years, 4 months ago) |
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First time working with agar. These are all from drips straight from syringes.
These are my best looking plates. i thought it’s weird that most the myceliums are kind of clear, but they’re starting to turn more solid white now so that’s cool I guess.
What’s with the one that’s kind of sectoring? I wasn’t expecting to see that yet.
Some obvious contams. Anything look particularly bad?
Any other advice?
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Lemgrub



Registered: 04/02/18
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Re: almost time for my first agar transfer i think. any advice? [Re: the dagda]
#27460724 - 09/08/21 05:04 PM (3 years, 4 months ago) |
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Just be sure to flame your scalpel red hot, maybe flame a little past the blade too. Cut away from the colony on blank agar to cool the blade. Be aware of what your hands are doing, don't let a hand go over the petri while open. Make smooth cuts.
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PussyFart
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Re: almost time for my first agar transfer i think. any advice? [Re: the dagda]
#27460759 - 09/08/21 05:30 PM (3 years, 4 months ago) |
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Quote:
the dagda said:
What’s with the one that’s kind of sectoring? I wasn’t expecting to see that yet.
Just a multitude of genetics in there. Some grow different, slower/faster thicker/thinner, etc. than others.
Take cuts from the fastest and healthiest growth.
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c10h12n2o
serial dilutor



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Re: almost time for my first agar transfer i think. any advice? [Re: PussyFart] 1
#27460981 - 09/08/21 08:32 PM (3 years, 4 months ago) |
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unless your plates are super dirty on the outside of the plate, all of those have pretty significant amounts of bacteria in them (the discolored blotches)
personally i would not transfer any of that, and would start fresh. you could probably get away with it, but thats way too much bacteria for me to be confident that you arent transferring some ride-along bacteria. bear in mind bacteria =/= wont work
i think the main problem is how long you waited to transfer, which let the bacteria grow large colonies
when working with spores i like to do serial streaks like in the guide in my sig, which gives you lots of colonies to choose from and makes it very easy to separate clean myc from contams.
when you allow your germination colonies to grow that large, especially from a spore syringe, they have way too much chance to intermingle with other stuff on the plate
for first transfers, i like to do them as soon as the colony is large enough to be easily recognized as cube myc (usually about half a grain of rice) and transfer several colonies to fresh plates, usually 3 per plate for the first transfer. this gives you lots to choose from for subsequent transfers to individual plates. at that point you start looking for sectoring etc
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the dagda
Stranger
Registered: 08/06/21
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Re: almost time for my first agar transfer i think. any advice? [Re: c10h12n2o]
#27461004 - 09/08/21 08:50 PM (3 years, 4 months ago) |
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oh man, that's a bummer to hear. I swear I'm cursed with too much patience.
I was waiting til I was sure they were cube myc. Since all but one were clear I wasn't sure yet.
What are the "large colonies" of bacteria that you see?
I see some streaks of discoloration from where the mss slid around during inoculation. Then there are pink dots on some of the plates. The one plate has a darker mold looking thing.
Thanks for the replies
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ichugwindex
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Re: almost time for my first agar transfer i think. any advice? [Re: the dagda] 1
#27461035 - 09/08/21 09:23 PM (3 years, 4 months ago) |
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For what its worth i think the agar on the left of your first pic looks like the best one out of these
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c10h12n2o
serial dilutor



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Re: almost time for my first agar transfer i think. any advice? [Re: ichugwindex] 1
#27461045 - 09/08/21 09:43 PM (3 years, 4 months ago) |
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that discoloration you are talking about looks like bacteria to me, water shouldnt change the color of agar like that
some types of bacteria pull the food coloring out exactly like that, its almost like they concentrate it, ive definitely seen it before on dirty plates that used food coloring. knowing the syringe had that much other junk in it, i wouldnt trust a colony that has grown out that far
that patience will come in handy! just start fresh and do a serial streak with a tiny amount of spore solution instead of letting it roll all over the plate.
you could keep transferring what you already have as well if you want but i would at least start a fresh germ plate as well
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C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide
"Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing."
"Convictions are more dangerous enemies of truth than lies"
― Friedrich Nietzsche
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the dagda
Stranger
Registered: 08/06/21
Posts: 30
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Re: almost time for my first agar transfer i think. any advice? [Re: c10h12n2o]
#27462345 - 09/09/21 08:02 PM (3 years, 4 months ago) |
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thanks! I'm still learning how to read plates.
I went ahead and did some transfers with what I had, if for no other reason than to see what happens.
Will do some new germ plates with a loop as shown too.
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sandman420
Saint PP



Registered: 06/17/04
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Re: almost time for my first agar transfer i think. any advice? [Re: the dagda]
#27462378 - 09/09/21 08:36 PM (3 years, 4 months ago) |
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These are all mold and a little bacteria. There is no good material.
Mold from the edge of the plate. Did you pour these in a still air box? I always got a lot of this type of edge contam when pouring in a SAB.
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the dagda
Stranger
Registered: 08/06/21
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Re: almost time for my first agar transfer i think. any advice? [Re: sandman420]
#27462427 - 09/09/21 09:29 PM (3 years, 4 months ago) |
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Are you saying all the mycelium is mold, not cube myc?
Yea, I used a SAB.
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sandman420
Saint PP



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Re: almost time for my first agar transfer i think. any advice? [Re: the dagda]
#27462649 - 09/10/21 04:58 AM (3 years, 4 months ago) |
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Yes that is what I am saying.
Pouring in a still air box is more likely to get contamination along the edges of the dish as it cools as you can see here. Several people will come tell you they never ever seent a problem with it but just look at it homey. Just look at it! You would be better off doing no-pour type agar. Such as pasty plates or what have you.
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