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OfflineFranknbeans
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Atl7 yellow contam
    #24258139 - 04/19/17 07:38 PM (7 years, 8 months ago)

First time Atl7 on agar (and new to agar < 100 plates).  First time I've seen something bright yellow.  Looked at forum and didn't spot similar pics.  Wondering what it is.  Rest of plate junky, first plate from Ms.


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Invisiblemynakedrat
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Re: Atl7 yellow contam [Re: Franknbeans]
    #24258173 - 04/19/17 07:48 PM (7 years, 8 months ago)

Did you sneeze on it?
Looks Like a bacteria

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Invisiblec10h12n2o
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Re: Atl7 yellow contam [Re: mynakedrat]
    #24258555 - 04/19/17 10:51 PM (7 years, 8 months ago)

looks pretty nasty...


id start over, and try this streaking technique with a wire loop



the colonies in zone 4 will be fairly easy to distinguish from contams


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

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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Invisiblemynakedrat
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Re: Atl7 yellow contam [Re: c10h12n2o]
    #24259371 - 04/20/17 10:25 AM (7 years, 8 months ago)

I'm gonna re do my plates tonight like that. I only scrubbed the surface once. I need to add more spore for shore

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OfflineMysticMoteToter
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Re: Atl7 yellow contam [Re: c10h12n2o]
    #24259495 - 04/20/17 11:45 AM (7 years, 8 months ago)

Quote:

c10h12n2o said:
looks pretty nasty...


id start over, and try this streaking technique with a wire loop



the colonies in zone 4 will be fairly easy to distinguish from contams



wouldn't opening the lid 5 times increase likely hood of contams 5x too? why not just dab the middle real quickly and be done with it?


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Invisiblemynakedrat
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Re: Atl7 yellow contam [Re: MysticMoteToter]
    #24259531 - 04/20/17 12:04 PM (7 years, 8 months ago)

It could, but you don't put anything over the plate except your sterilized loop, so if contams are a worry then, your SAB needs a good spray down, rinse off, etc.
And do more than one!

Edited by mynakedrat (04/20/17 04:51 PM)

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OfflineFranknbeans
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Re: Atl7 yellow contam [Re: mynakedrat]
    #24260025 - 04/20/17 04:37 PM (7 years, 8 months ago)

Thanks for feedback!.  Yes, I junked this one... Made 30, tossed this one and half of the rest ( transferred best 15 for next round).  I was surprised how weak they all looked relative to first plate cubes (where following same process of single drop from syringe yielded great results).  Will try suggested streaking when mix up new batch of agar(going to try mea with coffee)

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Invisiblec10h12n2o
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Re: Atl7 yellow contam [Re: Franknbeans] * 1
    #24260103 - 04/20/17 05:26 PM (7 years, 8 months ago)

yes, you are correct, it would increase the chance, but not enough to imact things, no more than any other time you open a plate

but this chance is already negligible when using proper technique in a SAB or FH, so might increase the contam chance from 0.001% to 0.005% or the like, certainly wouldnt make a difference in practice

i actually did my streaks wrong for the longest time, just randomly streakin, because i didnt really understand the point of what i was doing...

the way shown in that gif i like to share is basically a serial dilution, right on the plate. by streaking section 1, then flaming and dragging streaks from that to form zone 2, flaming and dragging streaks from zone 2 to make zone 3, and so on

each progressive zone gets thinner and thinner in terms of number of spores and contams present, and they will become more and more spread out, so that makes it quite easy to distinguish healthy myc from contams, and will have way fewer total strains present than a culture from a big blob of spores


why coffee btw? whats wrong with standard half strength MEA? it works fine in real labs, more than suitable for our projects. its basically the standard for mycology

i wouldnt experiment too much with the agar components, just get some ME and a bunch of A (i use 20g agar agar and 15g malt extract), and learn to make it well and consistently. no reason to reinvent the wheel: the standard(s) works just fine


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

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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OfflineFranknbeans
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Re: Atl7 yellow contam [Re: c10h12n2o]
    #24261097 - 04/21/17 05:01 AM (7 years, 8 months ago)

On the agar... This batch was potato not malt.  With the myc looking so thin and weak, was wondering if it had to do with agar choice.  Looking at some sclerotia posts, seemed most on mea.  There was a comment about coffee and possibly getting stones on agar (as way to more quickly identify better candidates for isolation and moving to grains).  So figured try mea, make some with 10%coffee/90% h20, some just mea.  See if atl7 happier and any diff with or without coffee.

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OfflineFranknbeans
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Re: Atl7 yellow contam [Re: Franknbeans]
    #24261104 - 04/21/17 05:08 AM (7 years, 8 months ago)

Oh.  Forgot to mention. THANK YOU for the written explanation on the streaking.  I watched the gif, but didn't fully appreciate the why and adv until I read your post.  Will no longer be dropping directly from syringe to agar and will be streaking instead.

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Invisiblec10h12n2o
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Re: Atl7 yellow contam [Re: Franknbeans]
    #24261817 - 04/21/17 01:12 PM (7 years, 8 months ago)

For sure buddy :highfive1:

I wish someone had explained it time that way a long time ago lol

It is a really useful technique and is Miles better then adding any additional water to your plate via syringe. That liquids mirrors things around makes it impossible to distinguish what's what

Additionally if you are working towards a limited or isolated culture, this technique can take about 100 transfers all of the total needed to get there. It's basically the same concept as a serial dilution in water but on the surface of the other instead

It's quite handy to be able to choose from hundreds of initial colonies


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

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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