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InvisibleLegionOfShroom
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Registered: 02/13/17
Posts: 235
Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Mateja]
    #27696120 - 03/15/22 09:47 AM (2 years, 14 days ago)

Thanks to everybody that has contributed to this thread, especially Mateja! This has been a great resource for me to learn from.

I made a 0.15% LME LC and inoculated with an agar wedge on 12/14. I stirred it a few times then left it sitting at room temp until 3/11.

On 3/11 I made another 0.15% LME and inoculated it with 6ml of LC from the jar that was made on 12/14. I’ve stirred the jar that was inoculated 3/11 once per day with a magnetic stir bar.

I’d like to use the 3/11 jar to inoculate 10+ grain bags, it would be my first time inoculating grain with an LC. I’ve had a great success rate inoculating bags with agar recently, but I’m interested in LC for faster colonization times.

I’m looking for feedback on the pictures that I’ve included at the bottom of this post. Also opinions on if it’s ok to move forward with this culture that sat on the shelf at room temp for about 90 days

I understand that it’s best to store your LC in the fridge once it’s colonized, I just forgot about it and made a mistake this time

Inoculated 12/14 (pint jar)
   


Inoculated 3/11 (quart jar)
 

I don’t have a control jar, after reviewing this thread I’ll probably start keeping one around

(note: the surface of the pint jar is more uneven than the quart jar, this distorts the light more and makes it harder to see objects through the liquid clearly during a turbidity test)


2% agar 2% LME test plate inoculated 3/11. This test plate was made immediately after inoculating the quart jar

   

The recovery in the petri dish seems a bit slow to me, but it looks like mycelium is starting to grow and I don’t see anything that looks like bacteria

All pictures in this post were taken today 3/15

Edited by LegionOfShroom (03/15/22 12:16 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: LegionOfShroom]
    #27696503 - 03/15/22 03:35 PM (2 years, 13 days ago)

:splooge:


That was a clean ass post in all regards, from the way you take pics to the LC recovering on plates, not to leantion your clean looking work space :thumbup:



Also you hardly even need control jars IMO when you know what to look for. An LC won't ever be 'slightly bacterial' it will either be completely turbid and you won't be able to see through the jar or it will remain crystal clear. Altho I'd advice everyone trying this out for the first time to use a control jar, especially those with a very limited experience and equipment unlike yourself :super:


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Cakes inside Water Tub

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InvisibleLegionOfShroom
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Mateja]
    #27696837 - 03/15/22 08:31 PM (2 years, 13 days ago)

Thanks so much, I’m very fortunate to have a dedicated space and good equipment to work with. I put a lot of care into my posts because I try to get the most out of these interactions. After posting I had second thoughts about if this was even the right place to post all of that stuff

This community has helped guide me through every significant milestone in my journey since I started my first PF Tek inside a SAB with some spore syringes back in 2017

I’m not always active here on a regular basis, but whenever I return it never fails to impress me how much knowledge is exchanged and how many talented individuals participate here

Thanks again Mateja and shoutout to everybody that helped me along the way

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InvisibleMateja
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: LegionOfShroom]
    #27696846 - 03/15/22 08:35 PM (2 years, 13 days ago)

And posters like yourself inspire me to put in more work into my own posts and to lvl up my equipment game for sure :thumbup:


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Cakes inside Water Tub

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InvisibleMateja
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Mateja]
    #27697118 - 03/16/22 12:10 AM (2 years, 13 days ago)

Quote:

justanotherhuman said:
Short version: does this rhizzy AF plate look bacterial to you? The culture is a clone of the pin in the center btw





You know what to do to test if it's bacterial or not :popcorn:


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Cakes inside Water Tub

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OfflineSpectreOfCommunism
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Mateja]
    #27699411 - 03/17/22 08:31 PM (2 years, 11 days ago)

Hey Mateja, is it really 0.05% ME by weight? That seems extremely low to me — for example 0.05% of 250ml water would be just 125mg


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Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus


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InvisibleMateja
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: SpectreOfCommunism]
    #27699490 - 03/17/22 09:25 PM (2 years, 11 days ago)

Weighing out 0.05% of LME is tricky enough just aim for inbetween 0.1 and 0.05 and you'll be good :super: idk about y'all's LME but mine comes out pretty dark even at 0.1 that's why I do 0.05% :shrug:


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Cakes inside Water Tub

Edited by Mateja (03/17/22 09:27 PM)

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Invisiblesandman420
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Mateja] * 1
    #27699765 - 03/18/22 05:34 AM (2 years, 11 days ago)

I do 10 times as much at 0.5% 18 grams to a gallon. Looks like this


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- Sandbag Tek - How To Sterilize Spawn Bags - All About Static Pressure / Pressure Drop for DIY Flow Hoods - Sandman's LC Tek-

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InvisibleSwabMarley
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: sandman420] * 1
    #27699807 - 03/18/22 06:41 AM (2 years, 11 days ago)

I normally go 0.2%. This is 0.6g in 300ml


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Offlinedyel
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: SwabMarley]
    #27699814 - 03/18/22 06:48 AM (2 years, 11 days ago)

I feel like dabbling in the arts of LC but don't have lme at hand, what is a good ratio for a honey or corn syrup recipe? 4%?

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InvisibleMateja
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: dyel]
    #27700180 - 03/18/22 01:05 PM (2 years, 11 days ago)

Idk why honey or corn syrup are supposed to be any different from LME? I'll be doing some 0.1% honey and 0.1% white sugar LC next time just to see what's up :takingnotes: I don't think anything unusual gonna be 'up' suga=suga


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Cakes inside Water Tub

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Offlinewonderousovoid
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Mateja]
    #27936122 - 09/05/22 08:31 PM (1 year, 6 months ago)

Should an agar wedge float or sink in an LC or does it matter?

I thought I read somewhere that floating was a bad sign for some reason but I can't seem to find much searching for LC and wedges floating vs sinking so I'm not sure where I read it or if I'm misrembering whatever I read

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Offlinenormalperson
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: wonderousovoid]
    #27937061 - 09/06/22 11:21 AM (1 year, 6 months ago)

large pieces of agar will float, and small pieces will sink. if left undisturbed the myc will colonize the surface and start pinning.

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OfflineLuminous7


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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: SwabMarley]
    #27947614 - 09/12/22 10:49 PM (1 year, 6 months ago)

Quote:

Myc Hunt said:
I normally go 0.2%. This is 0.6g in 300ml






Would it be essntial to add higher ratio of sugar if you want the LC jar to chill for a while? or is it possible the culture could stall out and die from lack of food?

I Have some LC's that are over a year old, stored at room temp and they looks alive and well. Ive had success with them to.

I was adding like 1-2 g per 500ml broth for the longer term ones, its a nice way to save cultures i beleive. They last incredibly long and you can just forget about them and have good LC at any time. I could imagine the shelf life would be alot longer if stored in the fridge. :peace:


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Invisiblehummingbird

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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Luminous7]
    #27947674 - 09/13/22 01:07 AM (1 year, 6 months ago)

I've been rocking LC since my first successful grow. That was over 10 years ago now...crazy, time flies. After a couple failed attempts at PF tek I actually had spores germinate in a vendor syringe, and that ended up going into a 4% corn syrup LC I made. Put it to wheat and spawned into a compost mix I used to use and got my first fruits. I actually had pretty good results going spore syringe to LC back then, but also a few failures here and there. I use less sugar and prints/swabs and agar now, but...

I've definitely noticed that bacterial LC gets cloudy really fast too. I've found that the green spore molds typically float to the top in a couple weeks or less and sporulate on the surface. I've been tricked by a black mold a couple times, even though something didn't seem quite right with the mycelium (extra whispy with odd clumps, and myc that stuck to the glass). Those cases, the myc grew way too fast on grains and turned black pretty quick.

I used to use metal lids with a hole sealed with red RTV silicone, but use plastic lids with a injection port and syringe filter and clear silicone ring seal now. I've found colonization speed is similar, but the cultures last longer without the filter. I had some LC still do well after 3 years or so at room temp back when I was using the metal lids with no filter. I'm trying out long term culture storage with .1% malt LC in capped syringes in the fridge now, I think it should work pretty well.

Ok, I'm done grandpa rambling now...cool thread, I love LC!

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