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Invisiblecoversall
إِنْ شَاءَ ٱللَهُ


Registered: 06/06/20
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: The Dalcassian]
    #27151513 - 01/16/21 03:28 PM (1 year, 10 days ago)

My first go at LC worked out pretty well and they were all in jars with unmodified lids. I did loosen the lids off after swirling.


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OfflineMLPismyOPSEC
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: The Dalcassian] * 1
    #27151518 - 01/16/21 03:31 PM (1 year, 10 days ago)

LC's do need some "GE" but the air left in the bottle is more than enough. I usually make 200mL of LC in a 250mL media bottle and that has been plenty enough air.


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Invisiblecoversall
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: MLPismyOPSEC] * 1
    #27151522 - 01/16/21 03:34 PM (1 year, 10 days ago)

Nice, I treated myself to a bunch of mini media bottles to make LC in and didn't want to mess up their lids.


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OfflineGrinchGrower
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: coversall]
    #27151530 - 01/16/21 03:43 PM (1 year, 10 days ago)

I did my first test using a trypsinizing flask w/ side arm and a 250ml media bottle.
The flask makes pouring really easy, but I wish I had a way to flame the neck/rim inside my SAB.


Think that is better suited for flow hood work though unless I want to blow myself up.
At least the caps are all graded for heavy sterilization in the PC...


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OfflineGrinchGrower
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: GrinchGrower]
    #27159394 - 01/20/21 01:49 PM (1 year, 6 days ago)

Need some advice...
I poured a splash of this to grain and a streak to agar five days ago.



I am not seeing any growth on either medium, and this flask did not show visible colonization until AFTER my pour/streak when I gave it a good shake and placed it back on the shelf.

Yesterday I noticed small roundish globules of mycelium floating around, so I put it on the mag plate for just a second to break it all up.

Today it looks like they consolidated into more of what I am used to seeing in other LCs here....

Is it possible to tell if the the mycelium looks healthy now, or if I picked up something nasty in my SAB during the first test?


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Invisiblemushhead
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: GrinchGrower]
    #27159648 - 01/20/21 03:51 PM (1 year, 6 days ago)

Quote:

GrinchGrower said:
I did my first test using a trypsinizing flask w/ side arm and a 250ml media bottle.
The flask makes pouring really easy, but I wish I had a way to flame the neck/rim inside my SAB.


Think that is better suited for flow hood work though unless I want to blow myself up.
At least the caps are all graded for heavy sterilization in the PC...




I like this.
:takingnotes: Added to the list of items needed for LC.


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OfflineLuminous7


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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: GrinchGrower]
    #27160191 - 01/20/21 09:13 PM (1 year, 5 days ago)

Quote:

GrinchGrower said:
Need some advice...
I poured a splash of this to grain and a streak to agar five days ago.



I am not seeing any growth on either medium, and this flask did not show visible colonization until AFTER my pour/streak when I gave it a good shake and placed it back on the shelf.

Yesterday I noticed small roundish globules of mycelium floating around, so I put it on the mag plate for just a second to break it all up.

Today it looks like they consolidated into more of what I am used to seeing in other LCs here....

Is it possible to tell if the the mycelium looks healthy now, or if I picked up something nasty in my SAB during the first test?




Thats basically why I dont bother testing mine. Just opens up another Vector of contamination.

I take all the precautions possible while making the LC and if it looks clean i use it.
So far this has been working for me.

Best of luck!


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


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Offlinekarri0n
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Mateja]
    #27293817 - 05/04/21 01:17 PM (8 months, 19 days ago)

Here is a mold LC, inoculated from clean GT mycelium via josex poke through a commercially produced SHIP:


After testing on grain:


I honestly didn't expect it to be mold, since it didn't float or sporulate.

In retrospect, I'd say I could see the following differences:

  • The broth had been cloudy and went clear. I thought it was bacterial, but it was also very dense grain water. I'm not sure at this time if it was sediment or bacteria.
  • colonization speed was faster than expected. In 12 hours the blob went from small little ball to 3/4 inch streak.
  • Some mycelium stuck to the glass and won't come off with a shake. I don't think I've ever seen mushroom mycelium do that.



I believe this failed LC is due to lid failure.

If you use plastic lids, always use gaskets, and don't buy the cheap thin red ones. Get the thicker clear silicone ones with a groove in them, from Amazon. The red gaskets are too thin and  difficult to position properly, and then they fall off when you screw the lid. After contaminating your jar several times to remove and replace lid to adjust the red gaskets, they then get brittle and fail in the pc. This is even worse if you are making your LC by opening the jar and transferring as opposed to using a SHIP. I wouldn't recommend opening an LC jar that used a thin red gasket at all. They are too hard to reseat in the sab.


--------------------
Micropore tape? More like myco-poor tape, right? HAR HAR


Edited by karri0n (05/04/21 01:18 PM)


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InvisibleMateja
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: karri0n]
    #27294702 - 05/04/21 11:25 PM (8 months, 18 days ago)

The whole point of this method is to use as crystal clear broth as possible so that you can easily determine when turbidity is occurring (Turbidity=bacterial colony present). So with that being said, to use a cloudy broth would totally defeat the purpose of this type of inspection.



I suggest using a 'control jar' which makes inspection much easier. A control jar is just a broth which hasn't been inoculated. Notice how easy it is to spot which LC's have gone bacterial!




Also for this specific method of inspection I'd advise inoculating with an agar wedge and what this does is allows you inspect the Cubensis culture as it's growing out from the agar wedge and this will give you an idea of how the cube culture looks and acts in the broth and this will enable you to easier spot potential mold colonies growing. And you have several types of white mold which will not sporulate visibly on neither plates nor in broths so don't bother looking for green sporulation as a means of detecting mold. In fact molds will always be able to blend in with healthy Cubensis mycelium and especially if you have a pretty far grown LC that only recently has been infected with mold. Gl


--------------------
Water Tub


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OfflineJohnS0N
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Mateja]
    #27608651 - 01/06/22 06:59 AM (20 days, 11 hours ago)

This thread is a really good reference point, both visual and contextual. Thanks for starting it and keeping it updated. :sogood:


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