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Offlinefreddyincognito
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Whats PE and what isnt?
    #24904960 - 01/11/18 07:59 AM (7 years, 6 days ago)

I recently got a spore syringe and decided to test it on 3 different pre-poured plates.
Whats myc and what isnt? Not sure what caused all this growth... the syringe was from a reputable vendor :shrug: But there is some small fuzzies, mostly massive transparent globs and some random other contams.



I thought the blobs for sure at first but now I don't know, there is some small white fluffy spots that look more like myc to me.

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Re: Whats PE and what isnt? [Re: freddyincognito]
    #24904987 - 01/11/18 08:24 AM (7 years, 6 days ago)

those blobs are bacteria. i would take the small fluffy stuff and keep the germ dish around to see if anything else pops up.


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Re: Whats PE and what isnt? [Re: Nobler Hino]
    #24906099 - 01/11/18 05:56 PM (7 years, 6 days ago)

:whathesaid:

From the first picture.


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Re: Whats PE and what isnt? [Re: freddyincognito]
    #24906105 - 01/11/18 05:59 PM (7 years, 6 days ago)

The first pic is myc. Second pic looks like all bacteria. 3rd pic the small circle of growth below the growth along the top edge looks like myc but it's hard to tell because the pic is blurry.


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Re: Whats PE and what isnt? [Re: FooMan]
    #24906152 - 01/11/18 06:29 PM (7 years, 6 days ago)

As you can see from all the bacteria its not enough to just "test" the syringe on agar. You need to transfer clean germination away from that bacteria to a new plate, get a clean culture to inoculate your grains with.


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Offlinefreddyincognito
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Re: Whats PE and what isnt? [Re: FooMan]
    #24906224 - 01/11/18 07:03 PM (7 years, 6 days ago)

Quote:

mushpunx said:
As you can see from all the bacteria its not enough to just "test" the syringe on agar. You need to transfer clean germination away from that bacteria to a new plate, get a clean culture to inoculate your grains with.




Yeah I know, that's why I made the post. Wasn't sure what to transfer!

Quote:

FooMan said:
The first pic is myc. Second pic looks like all bacteria. 3rd pic the small circle of growth below the growth along the top edge looks like myc but it's hard to tell because the pic is blurry.




I don't think the first or second pic have myc at all tbh... I think it's the brighter white fluffy spot on the 3rd pic. Much smaller than the bacteria so it's hard to see in the pic like you said. I'll try and transfer that.

EDIT: I'm gonna transfer the tiny myc and do a few new plates with the syringe to see if I can get any cleaner plates. Maybe my procedure was dirty?

Edited by freddyincognito (01/11/18 07:05 PM)

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Re: Whats PE and what isnt? [Re: freddyincognito]
    #24906261 - 01/11/18 07:19 PM (7 years, 6 days ago)

What was your procedure? :shrug:

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OfflinehamloafM
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Re: Whats PE and what isnt? [Re: freddyincognito]
    #24906333 - 01/11/18 07:43 PM (7 years, 6 days ago)

Quote:

freddyincognito said:
Quote:

mushpunx said:
As you can see from all the bacteria its not enough to just "test" the syringe on agar. You need to transfer clean germination away from that bacteria to a new plate, get a clean culture to inoculate your grains with.




Yeah I know, that's why I made the post. Wasn't sure what to transfer!

Quote:

FooMan said:
The first pic is myc. Second pic looks like all bacteria. 3rd pic the small circle of growth below the growth along the top edge looks like myc but it's hard to tell because the pic is blurry.




I don't think the first or second pic have myc at all tbh... I think it's the brighter white fluffy spot on the 3rd pic. Much smaller than the bacteria so it's hard to see in the pic like you said. I'll try and transfer that.

EDIT: I'm gonna transfer the tiny myc and do a few new plates with the syringe to see if I can get any cleaner plates. Maybe my procedure was dirty?



Nah, the first pic is the only one with myce.


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Re: Whats PE and what isnt? [Re: hamloaf]
    #24906337 - 01/11/18 07:46 PM (7 years, 6 days ago)

I think that's mycilium at 11 o'clock in the third pic about 1/4 the way up from center.

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OfflinehamloafM
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Re: Whats PE and what isnt? [Re: krypto2000]
    #24906356 - 01/11/18 07:52 PM (7 years, 6 days ago)

I think so too, but it's surrounded by bacteria.  The lonely myce culture in the first picture would be the safest place to transfer away from, but quickly before it joins with any bacteria cultures.


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Offlinefreddyincognito
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Re: Whats PE and what isnt? [Re: hamloaf]
    #24906378 - 01/11/18 08:00 PM (7 years, 6 days ago)

Quote:

krypto2000 said:
What was your procedure? :shrug:




Idk I was having trouble with the syringe at first so it exploded all over one of the plates :shrug: but I just lifted the lids under a SAB and used a flame sterilized needle. Same technique I've been doing so far on a bunch of other plates without any issues. Maybe it's just a bad syringe

Quote:

hamloaf said:
I think so too, but it's surrounded by bacteria.  The lonely myce culture in the first picture would be the safest place to transfer away from, but quickly before it joins with any bacteria cultures.




How can you guys tell that first pic is myc? It's not fuzzy at all and has the same gloss over like the bacteria. The white fluffy stuff you guys are talking about in the third pic is probably far enough from that bacteria that I can move it :thumbup: so I'll try that. Currently got some homemade petris cooling in the PC :biggrin:

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Re: Whats PE and what isnt? [Re: freddyincognito]
    #24906386 - 01/11/18 08:02 PM (7 years, 6 days ago)

Maybe the pictures aren't doing it justice from here but don't think it does.  It's a little tomemtose, but clearly has some rhizomorphic growth patterns from here.  The cultures in pictures two don't and three was surrounded.


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Offlinefreddyincognito
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Re: Whats PE and what isnt? [Re: hamloaf]
    #24906401 - 01/11/18 08:08 PM (7 years, 6 days ago)

I see the rhizomorphic growth under the gloss actually, it's different than the bacteria. Maybe I will try and transfer that one and see what happens.

Edit: heres a better pic of the same spot


Edited by freddyincognito (01/11/18 08:11 PM)

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Re: Whats PE and what isnt? [Re: freddyincognito]
    #24906420 - 01/11/18 08:17 PM (7 years, 6 days ago)

Ah, can see the shine now.  Thanks for the better pic.  Doesn't look like myce.  Sorry, brother.


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Re: Whats PE and what isnt? [Re: hamloaf]
    #24906430 - 01/11/18 08:22 PM (7 years, 6 days ago)

That tiny dot at 11 o clock underneath that bacteria culture that's touching the wall of that plate is myce.  Transfer it soon if you wish to attempt to clean that plate.


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Offlinefreddyincognito
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Re: Whats PE and what isnt? [Re: hamloaf]
    #24906438 - 01/11/18 08:24 PM (7 years, 6 days ago)

I think that's my best bet. Damn, I wonder what happened with this syringe. Hopefully I'll pull a clean plate out of this.

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Re: Whats PE and what isnt? [Re: freddyincognito]
    #24906662 - 01/11/18 10:15 PM (7 years, 6 days ago)

also if i may add, its a good idea to make several plates when your starting from spore or tissue cloning for that matter. instead of three plates, make 5 or 6.


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Re: Whats PE and what isnt? [Re: freddyincognito]
    #24907041 - 01/12/18 04:39 AM (7 years, 5 days ago)

Quote:

freddyincognito said:
I think that's my best bet. Damn, I wonder what happened with this syringe. Hopefully I'll pull a clean plate out of this.




Did you unwrap those plates just for the photo or did you leave them like that after inoculation? Or is the edge wrapped but cant see in the photo?

Next time you inoculate from syringe, place a single drop onto the agar and then use a sterilized inoculating loop to swipe the spores across the plate in a zig zag pattern :thumbup:
Otherwise the spore solution will roll around the plate. With a swipe you should have a good idea where to expect germination


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Offlinefreddyincognito
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Re: Whats PE and what isnt? [Re: mushpunx]
    #24907105 - 01/12/18 05:52 AM (7 years, 5 days ago)

Yeah I'll make more plates I was running low.

Quote:

mushpunx said:
Quote:

freddyincognito said:
I think that's my best bet. Damn, I wonder what happened with this syringe. Hopefully I'll pull a clean plate out of this.




Did you unwrap those plates just for the photo or did you leave them like that after inoculation? Or is the edge wrapped but cant see in the photo?

Next time you inoculate from syringe, place a single drop onto the agar and then use a sterilized inoculating loop to swipe the spores across the plate in a zig zag pattern :thumbup:
Otherwise the spore solution will roll around the plate. With a swipe you should have a good idea where to expect germination




Are you supposed to wrap those plates in something? I left them just as they are after inoculating.

Oh I'd never heard of that technique with the inoculation loop :thumbup: will try that. Is a bent safety pin good enough as a homemade inoc loop or should it be the real thing?

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Re: Whats PE and what isnt? [Re: freddyincognito]
    #24907117 - 01/12/18 06:05 AM (7 years, 5 days ago)

Well that could be your main problem right there!



After inoculation plates should be wrapped tightly along the edge with Parafilm (a stretchy, wax coated lab tape specifically for wrapping plates + flasks), or plastic cling wrap (saw  a 1" section off the end of a roll to make it easy).
Above shows a photo of one of each in that order.

Un inoculated plates should either be stored in their sleeve, or in fresh (fresh are sterile) ziplock bags, or wrapped. Don't let the top of the plate come off.

Are you working in a still air box (SAB)?

If you are going to continue working with agar, and I suggest you do, really do your reading on sterile technique and agar work first!

:thumbup:


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