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InvisibleD3_Myc
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Community Experiments and Projects Thread πŸ”¬ * 8
    #27003791 - 10/25/20 10:40 PM (3 years, 3 months ago)

Welcome To the Community Experiments and Projects Thread


This is a place where we can share experiments and projects we are currently working on and update in a singular post.

All are welcome to participate and the rules are simple.


  • Title your post

  • State your intentions: be it an experimental procedure, design or testing out new recipes and concepts.

  • Formulate a hypothesis: A prediction of what will be found at the outcome of your project work.

  • Share your experiments to test your hypothesis

  • Continue to update your original post: Utilize the edit button to update your OP with the results and conclusions of your experiments or project work.

    Side conversations are acceptable and are to be expected but for the sake of keeping the thread clean, organized and searchable keep your experiments and project work contained within your original post.

  • Have fun: Keep in mind not everyone is a scientist or have backgrounds in mycology. Realize that some experiments may be flawed or lack proper controls. You can offer suggestions to the OP's to advance their work but try not to flame, harass or belittle them in regard to their attempted experiments.

  • Experiments can be open to the public: Other members of the community may opt in to help test your hypothesis with their own experiments and share their data with you.


I will post the links to your experiments below with your thread title so they are easily accessed by others.

I look forward to seeing everyone's work and sharing our progress with one another.

Community Projects


The Potential Negative Effects of Erythrosine (Red#3) on Mycelium. A Comparison of Growth on Assorted Dyed Agar Mediums

Influence of agar stiffness and nutrition on mcyelium


Edited by D3_Myc (11/02/20 09:24 PM)


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InvisibleD3_Myc
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Re: Community Experiments and Projects Thread [Re: D3_Myc] * 5
    #27003803 - 10/25/20 10:44 PM (3 years, 3 months ago)

The Potential Negative Effects of Erythrosine (Red#3) on Mycelium. A Comparison of Growth on Assorted Dyed Agar Mediums

Background: Anecdotal reports suggest that mycelia grown on agar containing red#3 a artificial  dye called Erythrosine may cause irregular, sporadic or stunted growth of mycelia. Erythrosine is an artificial red (cherry-pink) food coloring made from coal tar. It is an organic compound containing iodine and sodium. Limited research suggests that Erythrosine may have natural fungicidal properties.

The intent of this experiment will be to test out growth patterns of mycelium on agar dyed with various colors of food coloring to determine if any have benefits over the others or perhaps hinder growth.

Hypothesis: The agar plates containing erythrosin (red#3) or in this case Wilton neon gel magenta food coloring will have a negative impact on mycelium growth.

Experiment:

All agar recipes consist of 20g agar agar, 12.5g Light malt extract and 1000ml of distilled water. Dyes selected are Wilton Neon Gel "Magenta", "Teal" and organic charcoal powder. ( I realize a control plate with no coloring would have been beneficial to the experiment but time did not allot having some poured in time. Hopes is that the charcoal powder (black plates) are on a color media that shouldn't be providing any additional nutrients in the form of corn sugars or other ingredients that are found in the gel food coloring.

Sample #1



Transfers where taken on 10-23 from equally healthy looking growth using a custom agar punch tool keeping transferred samples uniform in size and shape.





Sample #2



Again transfers taken on 10-23



All test subjects are stored in a manor that they receive similar light and rotated every two days to ensure equal lighting and heating conditions are obtained.



Update 10-26


Sample #1



Sample #2



Update 10-27-20


Fairly easy to tell there’s there’s a difference in growth rate between the red plates and other dishes.

Sample 1



Sample 2




Update 10-29-20

I think it's fairly evident that there's stunted maybe even halted growth on the plates containing Erythrosine (Red #3) Compared to the other plates.

Sample 1



Sample 2




I don't see any noticeable differences between the blue or black plates in growth does anyone else really?

For my next samples I will pour the same nute levels but no dye, purple (which contains both red #40 and red #3 ) orange, blue and black and do a comparison.

Update 10-31 Happy Halloween!! πŸ‘» πŸŽƒ


This is my final update with this batch of samples. I’ll let you draw your own conclusions based on the photographic evidence provided



For my next round I’m expanding to include

  • Purple

  • orange

  • No coloring


Stay tuned

Unofficial final final pic.. lol




Edited by D3_Myc (11/06/20 11:01 PM)


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InvisibleModularMind
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Re: Community Experiments and Projects Thread [Re: D3_Myc] * 1
    #27003854 - 10/25/20 11:28 PM (3 years, 3 months ago)

:havesomescience:

:superscience:


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InvisibleDoctor Mario
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Re: Community Experiments and Projects Thread [Re: ModularMind] * 1
    #27005487 - 10/26/20 10:02 PM (3 years, 3 months ago)

Hell yes, thank you for doing this.


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OfflineSingularFusion
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Re: Community Experiments and Projects Thread [Re: Doctor Mario]
    #27006097 - 10/27/20 11:14 AM (3 years, 3 months ago)

:takingnotes:


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Re: Community Experiments and Projects Thread [Re: SingularFusion]
    #27006171 - 10/27/20 11:42 AM (3 years, 3 months ago)

Well this explains a lot...I randomly have lackluster plates for seemingly no reason and the only food coloring I've ever used is red. I'm gonna be getting some new colors today and see what happens on my end.

I would be interested to find out if the red#3 affects further transfers off of it or would the mycelium bounce back if transferred to say orange or black.


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Invisibledfwerydfhg
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Re: Community Experiments and Projects Thread [Re: pureshrooming]
    #27006775 - 10/27/20 05:24 PM (3 years, 3 months ago)

following


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Re: Community Experiments and Projects Thread [Re: dfwerydfhg] * 1
    #27006903 - 10/27/20 06:35 PM (3 years, 3 months ago)

Red never gave me issues:shrug:




I only use a few drops tho.


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Re: Community Experiments and Projects Thread [Re: mushboy]
    #27006911 - 10/27/20 06:38 PM (3 years, 3 months ago)

i bet if you took a transfer from the red plate to a blue plate it would outpace the donor plate in a few days.


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Re: Community Experiments and Projects Thread [Re: JHOVA]
    #27006921 - 10/27/20 06:42 PM (3 years, 3 months ago)

Lile, I've transfer between agar colors all the time. Without any thought or order or noticable effect:shrug:


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InvisibleD3_Myc
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Re: Community Experiments and Projects Thread [Re: mushboy]
    #27006981 - 10/27/20 07:10 PM (3 years, 3 months ago)

What brand you using? Maybe different dye used


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Re: Community Experiments and Projects Thread [Re: D3_Myc]
    #27006992 - 10/27/20 07:14 PM (3 years, 3 months ago)

McCormick. Basic shit in the grocers isle.

Isnt food coloring sugar? I use 2drops for 1000ml. Maybe making the color too deep is adding too much sugar?:strokebeard2:

Shit: google says most food coloring is made from oils like petroleum??

Quote:


Much red food coloring, known as carmine or cochineal, is made from a white insect that exudes a bright red color when it is crushed.





:justdontknow:


Edited by mushboy (10/27/20 07:19 PM)


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InvisibleNobler Hino
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Re: Community Experiments and Projects Thread [Re: mushboy]
    #27007149 - 10/27/20 08:35 PM (3 years, 3 months ago)

I use red dye all the time. For a couple years now. I haven't noticed any issues but I only used one drop per 500ml. Plates will even turn back to normal agar color when it's fully colonized.

As for the brand, I can't remember what I have.Cheapest option online I'm sure.


Edited by Nobler Hino (10/27/20 08:40 PM)


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InvisibleD3_Myc
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Re: Community Experiments and Projects Thread [Re: pureshrooming]
    #27007906 - 10/28/20 09:01 AM (3 years, 2 months ago)

Quote:

pureshrooming said:
Well this explains a lot...I randomly have lackluster plates for seemingly no reason and the only food coloring I've ever used is red. I'm gonna be getting some new colors today and see what happens on my end.

I would be interested to find out if the red#3 affects further transfers off of it or would the mycelium bounce back if transferred to say orange or black.




I can try that after they have grown out a bit. I would like to test it against a few other brands as well. Perhaps the wilton gel has a higher concentration of erythrosin than say the non gel coloring. Looking at Mc Cormicks red dye looks like they use both Red 40 and Red #3 . They also have a natural dye that uses beet juice for the red coloring. Just depends on the kind you buy. In this case I used 1 drop per 100ml of the wilton neon gel


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Re: Community Experiments and Projects Thread [Re: D3_Myc]
    #27008721 - 10/28/20 03:58 PM (3 years, 2 months ago)

This is a great experiment! I am interested if other dyes show similar antifungal properties.

I just made PDA with some of this colored agar I haven't used before.


It contains Red #40 and so far is doing well. Actually outperforming my MEA formula...


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InvisibleD3_Myc
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Re: Community Experiments and Projects Thread [Re: mushboy]
    #27008782 - 10/28/20 04:35 PM (3 years, 2 months ago)

Quote:

mushboy said:
McCormick. Basic shit in the grocers isle.

Isnt food coloring sugar? I use 2drops for 1000ml. Maybe making the color too deep is adding too much sugar?:strokebeard2:

Shit: google says most food coloring is made from oils like petroleum??

Quote:


Much red food coloring, known as carmine or cochineal, is made from a white insect that exudes a bright red color when it is crushed.





:justdontknow:




It’s amazing some of the stuff the food and drug industry gets away with. I remember watching a β€œhow it’s made” type deal on how cod fish sperm is used as a binder in lipstick. They spent a while interviewing this poor dude who’s sole industry role is hovering over a conveyor belt extracting fish sperm all day... makes one appreciate a few of their life choices. Unless they are into that sort of thing lol


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Re: Community Experiments and Projects Thread [Re: D3_Myc]
    #27009024 - 10/28/20 06:55 PM (3 years, 2 months ago)

I just picked up an assortment of colors in gel and regular so I'll be whipping some plates up tonight. Honestly tho that black charcoal seems like the best option to not have to worry about any random ingredients.


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InvisibleDoctor Mario
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Re: Community Experiments and Projects Thread [Re: pureshrooming]
    #27011074 - 10/29/20 08:05 PM (3 years, 2 months ago)

I knew I wasn't trippin. I also used q drop of the red food coloring per 100mL water. Its like the shit just wouldn't expand and when it finally did, it grew underneath the agar and moved very slowly probably because it had to fight to get through the agar.


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Re: Community Experiments and Projects Thread [Re: Doctor Mario]
    #27012577 - 10/30/20 06:12 PM (3 years, 2 months ago)

Great idea love to see what comes of this thread


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InvisibleD3_Myc
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Re: Community Experiments and Projects Thread [Re: MycFunk3D]
    #27014901 - 10/31/20 11:47 PM (3 years, 2 months ago)

I've updated the coloring experiment and have poured some additional plates to add to it. Anyone else got any fun stuff they are working on they would like to share?


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Re: Community Experiments and Projects Thread [Re: D3_Myc] * 6
    #27017287 - 11/02/20 11:12 AM (3 years, 2 months ago)

Influence of agar stiffness and nutrition on mcyelium


Background:
I am currently unhappy with my current agar recipe. Tomentose mycelium and 'translucent' growth is more prevalent than rhyzomorphic. Also there has been an ongoing discussion since the beginning of time what actually causes mycelium on agar plates to grow either rhyzomorphic or tomentose, complicated by the fact that the mycelium seemingly can change from one form into the other by transferring it into a new plate even when using the same agar recipe. The common opinion seems to be that a lower concentration of nutrients will favor rhyzomorphic growth, while a higher amount tomentose. One of the arguments is that in plates with less nutrition the mycelium will grow quicker and more vigorous in search of food. 
This scenario has been challenged in the past by investigations like these:
[Rizo & Tomentose] vs [Weak & Strong Plates] vs [Soft & Hard] pucks
where the opposite was reported: The higher the amount of nutrients, the more rhyzomorphic and faster the culture was growing.
Rhyzomorphic growth is regularly reported when a culture hits the edge of the plate and growths up the side of the petri dish, where it often changes from previously tomentose on agar to rhyzomorphic on the plastic. This observation can support the less nutrition=more rhyzomorphic theory. On the other hand, one often observes that a culture when transferred to grain shows strong rhyzomorphic growth, even when it did not on agar, which would support the opposite more nutrition=more rhyzomorphic theory. Motivated by these issues and conflicting reports & theories I started my own investigation with different agar recipes.

Hypothesis:
My hypothesis is actually different from the two common ones: I propose that rhyzomorphic growth is mostly triggered by the stiffness of agar, i.e. how far it can penetrate the medium. The softer the agar, the more it will show tomentose growth, while the stiffer the agar, the more it will be rhyzomorphic. My theory is that the tomentose growth is some kind of overlay that develops on the surface when the mycelium is actually growing through a medium like agar. If penetration is reduced by an increased hardness of the agar, it stays mostly on the surface and forms rhyzomorphic growth.

This theory was inspired by an observation when I scratched the surface of different mycelium. Below the tomentose myc below the agar surface there was a milky/dull layer where the myeclium had obviously grown into the agar. This layer could often be less than 1 milimeter, but sometimes stretched through the full width of the agar puck. To exclude that this layer could be some kind of bacterial contamination I transferred a tiny piece from this region below the surface and it grew out into healthy normal looking mycelium.

On the other hand, for most rhyzomorphic mycelium such a layer was almost nonexistent. This theory would also be consistent with the reports of agar climbing the sides of the petri dish and rhyzomorphic growth on grains, since both allow for less penetration than agar. Low nutrients could also discourage the mycelium from growing into the agar, essentially mimicking this effect.
The observation in the thread above (more nutrients=more rhyzo) are still challening. Maybe the increase in nutrients also led to a stiffer agar.
Therefore, the time has come for another systematic investigation.:takingnotes:

Experiment:
The experiment consists of using different agar recipes with different amount of nutrients and stiffness. I will be using grain water agar that can be diluted to change the amounts of nutrients.
The current samples are:
  • 100% grain water , 2.0% agar
  • 100% grain water , 2.5% agar
  • 100% grain water , 3.0% agar
  • 50% grain water , 2.0% agar
  • 50% grain water , 2.5% agar
  • 50% grain water , 3.0% agar
  • 25% grain water , 2.0% agar



After pouring plates hey will be inocculated with a multispore culture and a clone (both cubensis).
Right now I am PC'ing the agar, so the experiment has just started.

*** Update 10.Nov.2020 ***

It's now day 4 since the first batch of plates has been inocculated. I took small transfers from a MS x7x plate which looked clean an showed neither pronounced rhizomorphic or tomentose growth. So far no clear differentiation is visible between the different plates. Will update in a few days again.


I started another batch with a GT clone, and a Natalensis/Natal MS, which are two days behind, so they will also show up in the next update.


Edited by sporecap (11/10/20 01:10 PM)


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InvisibleD3_Myc
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Re: Community Experiments and Projects Thread [Re: sporecap]
    #27018372 - 11/02/20 09:31 PM (3 years, 2 months ago)

Awesome thanks for sharing.

Another viewpoint with same conclusion is increased agar rigidity means less nutrients are bio-available as the mycelium has a harder time penetrating the media in search of food and instead spreads out seeking the meager amount of nutrients that are made bio-available along the mediums surface.

I look forward to seeing your results.


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Re: Community Experiments and Projects Thread [Re: sporecap]
    #27018399 - 11/02/20 09:50 PM (3 years, 2 months ago)

:takingnotes:


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Re: Community Experiments and Projects Thread [Re: SingularFusion] * 1
    #27025849 - 11/06/20 10:48 PM (3 years, 2 months ago)

So interesting, thanks for sharing!


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Re: Community Experiments and Projects Thread [Re: Sandro]
    #27026808 - 11/07/20 01:00 PM (3 years, 2 months ago)



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Re: Community Experiments and Projects Thread [Re: psillyboy]
    #27031623 - 11/10/20 01:11 PM (3 years, 2 months ago)

Updated post #27017287


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Re: Community Experiments and Projects Thread [Re: sporecap]
    #27031853 - 11/10/20 03:37 PM (3 years, 2 months ago)

Nice experiment


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Re: Community Experiments and Projects Thread [Re: sporecap]
    #27037098 - 11/13/20 01:38 PM (3 years, 2 months ago)

Quote:

sporecap said:


It's now day 4 since the first batch of plates has been inocculated. I took small transfers from a MS x7x plate which looked clean an showed neither pronounced rhizomorphic or tomentose growth. So far no clear differentiation is visible between the different plates. Will update in a few days again.


I started another batch with a GT clone, and a Natalensis/Natal MS, which are two days behind, so they will also show up in the next update.




seems like the clone is a better candidate for a comparison as transfers from a multispore plate are going to give you pretty random samples.

i have been led to believe the opposite about rhizo growth stemming from more nutrients.  Been cutting back nutes batch by batch in my agar going for that rhizo growth.

very interested to see the progress.

d3: did you ever calrify the exact amnount of charcoal in your agar?


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Re: Community Experiments and Projects Thread [Re: nmd_myco]
    #27037318 - 11/13/20 04:01 PM (3 years, 2 months ago)

I don’t believe it did, I typically don’t measure it but ~1g charcoal powder to 100ml is roughly what I average.


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Re: Community Experiments and Projects Thread [Re: D3_Myc]
    #27087614 - 12/13/20 05:13 PM (3 years, 1 month ago)

great thread buddy!!

i been using the exact same food coloring as D3monic and noticed the same stunted growth on the magenta/red/purple mixes

oddly enough, it doesnt always happen, but it does happen often enough to notice the pattern. amazing cultures would sometimes just look like shit for no reason, and pan myc would often just stop growing halfway through the plate

imma give this charcoal stuff a try, looks sexy. D3monic is a trendsetter with agar fashion haha


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"Convictions are more dangerous enemies of truth than lies"
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Re: Community Experiments and Projects Thread [Re: c10h12n2o]
    #27087701 - 12/13/20 06:07 PM (3 years, 1 month ago)

Nice seeing more confirmation of this.  Since I stopped using red, I stopped having any of the weird issues.  Blue and black ftw.  I was looking back through old notes the other day and came upon the first time I had ever had a coloring related issue.  At one point I thought fuck it and used a drop of black tattoo ink (I am aware they aren't the same thing, for the record...) because I was bored (had a bottle I was unlikely to use again), and it made for great coloring and my plates showed totally normal growth.  Me being a cheap ass, I was content to use it up.  Did that twice all was well.  The third time I accidentally squirted like 3 or 4 drops in there and bam: weak growth,  translucent growth,  etc across a couple dozen plates.  Transferred to a new round with just a drop and everything was kosher again.  I genuinely think it's a threshold issue,  but it definitely seems to be a thing.  Getting used to all blue plates and now I kind of dig it.  1-2 drops for 500ml and no issues at all.  I went once to test the threshold theory with blue and growth looked weaker and was slower, but with red it would have been potato.


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Community Experiments and Projects Thread [Re: alaskappalachian]
    #27087743 - 12/13/20 06:31 PM (3 years, 1 month ago)

Multi Isolate cocktail
Almost identical to mixing MS from multiple varieties with similar outcome just with an extremely limited gene pool.
Not a crossing attempt.No reason to believe it will increase production in any way. I just really want to see all them crossing swords.
My plan is to add a quarter wedge from each culture into one pint. Then G2G with that pint. Would anyone take a different approach? I urge anyone with 3 or more distinct isolates/clones to give it a try. It does not need to be PE varieties but they should be distinct ISO/clones.
At the moment I have these three cocks that are distinct enough.
Ape clone,Melmek clone,Penis envy clone (Boner cloner). I have Lpeu swabs so eventually this will be in the mix but a clone is important so it will take time.Also have my fingers crossed that ill get my hands on that Toc Cock.wink wink :blush:
I have toyed with the idea before but untimely decided it was a waste of time. Now i will just do it for the hell of it.

-Pointless waste of time.
-Potential way to get diverse yet controlled grows.
-Decreased efficiency from known cultures.
What are your thoughts?

A small list of "projects Ill totally do one day


Edited by bw86 (12/13/20 06:49 PM)


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Registered: 01/18/21
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Re: Community Experiments and Projects Thread πŸ”¬ [Re: D3_Myc]
    #27188029 - 02/05/21 01:17 PM (2 years, 11 months ago)

This needs to be a sticky or something, noob here started w red agar last year and was wondering wtf ingredient was killing my stuff (yeah we actually did manage to get a resistant collection going, its slow)


PastyWhite uses it in his agar tek and that's the noob go-to, it at least needs to be noted there in bold somewhere


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yeah keep telling yourself that's voluntary


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