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Offlinechowyunfat
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Suspicious stuff in agar after 48hrs
    #27253479 - 03/15/21 07:07 AM (2 years, 10 months ago)

as agar beginner I must ask, is this "normal" or is it already bacterial after 48hrs?

agar is inoculated from syringe, black clump came out of syringe...seems like all water from syringe that dropped off turned out to suspicious white stuff?
All plates inoculated from syringe have this.
Plates I inoculated from print look fine.


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Offlinepesa
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Re: Suspicious stuff in agar after 48hrs [Re: chowyunfat]
    #27253501 - 03/15/21 07:24 AM (2 years, 10 months ago)

that's not normal its bacteria, you might get some mycelium out of it but bacteria will keep riding it. better try this method to get bacteria free mycelium.

https://www.shroomery.org/forums/showflat.php/Number/24806569


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Offlinekanemush
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Re: Suspicious stuff in agar after 48hrs [Re: chowyunfat]
    #27253502 - 03/15/21 07:25 AM (2 years, 10 months ago)

yup those look screwed sorry to say unless you shot a shit load of spores on them.


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Offlinechowyunfat
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Re: Suspicious stuff in agar after 48hrs [Re: kanemush]
    #27253509 - 03/15/21 07:34 AM (2 years, 10 months ago)

Quote:

pesa said:
that's not normal its bacteria, you might get some mycelium out of it but bacteria will keep riding it. better try this method to get bacteria free mycelium.

https://www.shroomery.org/forums/showflat.php/Number/24806569



Quote:

kanemush said:
yup those look screwed sorry to say unless you shot a shit load of spores on them.




you think it's done? no chance it will develop a bit of myc so I can clone the healthy part to another plate?

thanks Pesa, will check the link


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Offlinepesa
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Re: Suspicious stuff in agar after 48hrs [Re: chowyunfat]
    #27253520 - 03/15/21 07:41 AM (2 years, 10 months ago)

it will take a lot of time for mycelium to escape bacteria and it will look more weird like this



or bit different depending on mycelium and amount of bacteria but bacteria will keep riding it. just use josex method it works every time and you will get clean mycelium.

the pic above i kept transferring after about 5 transfers later, i realize its useless and your plates are very heavily bacterial wait for a week to see if mycelium can escape otherwise sorry to say plate is pretty much done.


Edited by pesa (03/15/21 07:43 AM)


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Offlinechowyunfat
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Re: Suspicious stuff in agar after 48hrs [Re: pesa]
    #27253537 - 03/15/21 07:50 AM (2 years, 10 months ago)

Quote:

pesa said:
it will take a lot of time for mycelium to escape bacteria and it will look more weird like this



or bit different depending on mycelium and amount of bacteria but bacteria will keep riding it. just use josex method it works every time and you will get clean mycelium.

the pic above i kept transferring after about 5 transfers later, i realize its useless and your plates are very heavily bacterial wait for a week to see if mycelium can escape otherwise sorry to say plate is pretty much done.



huh, he calls it noobish but it looks like demanding procedure... must read again, not sure if I understood it all, he takes a tiny bit and inoculate a bit of substrate, which then produces a bit of healthy myc, and then you isolate that huh?

there is no way I can figure out if the syringe is f-d up (ie person who provided it didn't do a good job) or my operation was not sterile enough... but if plates from print turn out fine I can maybe suspect it's not me :laugh:


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OfflineProfessor X
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Re: Suspicious stuff in agar after 48hrs [Re: chowyunfat]
    #27253546 - 03/15/21 07:55 AM (2 years, 10 months ago)

I have a bacterial culture that started from a syringe. Ive transfered grain of salt size transfers twice and still have a pour ringed puffball. I am going to go to BRF cake next then back to agar.


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

My no pour Petri TEK - https://www.shroomery.org/forums/showflat.php/Number/27252059/page/1


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InvisibleBenson
The Kidd ⭐


Registered: 09/29/20
Posts: 837
Re: Suspicious stuff in agar after 48hrs [Re: Professor X]
    #27253629 - 03/15/21 08:46 AM (2 years, 10 months ago)

Read through this thread, a guy recently took a really nasty spore syringe and turned it into beautiful myc by using some cool teks. I'm sure you could replicate the process!

https://www.shroomery.org/forums/showflat.php/Number/27186099


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Offlinepesa
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Re: Suspicious stuff in agar after 48hrs [Re: chowyunfat]
    #27253636 - 03/15/21 08:49 AM (2 years, 10 months ago)

he calls it noobish since he is using pf tek and making small little pucks of pf cakes.

you said from SPORES its a clean growth that tells a lot about sterilization method you using since with spores every thing is going well and take pics of spore plates and show the growth and show pics of plates with bacteria either get a new syringe or get your money back from person who sold you this syringe.

also what tek you are using for agar? and did you do it in front of a flow hood or Still air box?


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Offlinechowyunfat
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Re: Suspicious stuff in agar after 48hrs [Re: pesa]
    #27253824 - 03/15/21 11:14 AM (2 years, 10 months ago)

Quote:

pesa said:
he calls it noobish since he is using pf tek and making small little pucks of pf cakes.

you said from SPORES its a clean growth that tells a lot about sterilization method you using since with spores every thing is going well and take pics of spore plates and show the growth and show pics of plates with bacteria either get a new syringe or get your money back from person who sold you this syringe.

also what tek you are using for agar? and did you do it in front of a flow hood or Still air box?



I had some pre made malt extract agar mix which I pc-ed at 15 PSI (I have glass plates, so I pc-ed them too) and then inoculated in still air box.
We'll see how plates inoculated with spore print from the same round will do!


while we talk about agar maybe someone can advise,
last time I experimented with it I had problems with
my knife leaving black burn marks after flame sterilizing on agar while cutting it.
Any way to avoid that? I use surgical knife and surgical blades.
I tried flame sterilizing with small alcohol lamp burner and regular lighter, seems like knife always goes black and leave burn marks ...


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Offlinepesa
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Re: Suspicious stuff in agar after 48hrs [Re: chowyunfat]
    #27253904 - 03/15/21 12:36 PM (2 years, 10 months ago)

if you can get a new syringe or money back that would be nice,

i use borosilicate glass petri dish. after cleaning and washing with anti bacterial soap pc them in glass jars never got a problem. just making sure all petri dish are dry before going to pc.

i use surgical knife never had a problem with those black marks usually mycelium end up either eating it or cover it, as long as your blade is glowing red hot you are fine.


this a ksss mycelium. they are too wispy because i was using very low low nutrient agar



you can see the cut mark at 6 o clock with bit of black marks left from blade

after transfer on very rich nutrient agar. i use PDA(potato dextrose agar) there are few bits from potato's in agar never bother me or gave problem



its going well as long as you make sure blade is glowing red hot and cool it on agar it will be okay.

sorry i use potato cam 3000 for all my pics


Edited by pesa (03/15/21 12:39 PM)


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