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InvisibleAtomsplit
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Mold on agar/ First transfer selecting mycelium spore print to agar
    #27329636 - 05/30/21 10:46 PM (15 days, 6 hours ago)

I've been having a string of failures trying to get a grow going.

I think I got mold growing on some spore print to agar 6 days ago.



Any opinions would be appreciated.
Thanks in advance.


Edited by Atomsplit (06/06/21 02:57 PM)


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InvisibleGoatriderS
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Re: Mold on agar? [Re: Atomsplit]
    #27329645 - 05/30/21 10:55 PM (15 days, 6 hours ago)

I`d wait a few days more to have something to transfer.
Germination plates usually look ugly :wink:

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OfflineCrackatoaS
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Re: Mold on agar? [Re: Goatrider]
    #27329650 - 05/30/21 11:02 PM (15 days, 6 hours ago)

Are those places you put spores? If so :whathesaid:


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Re: Mold on agar? [Re: Crackatoa]
    #27329666 - 05/30/21 11:11 PM (15 days, 5 hours ago)

Yes for the most part. I zigzaged swiped the agar and looked at
the thin lines pre and post growth.

I wouldn't doubt a satellite mold growth though on the orange plate though.

Maybe I rubbed off most of the spores at the beginning of the swipe? This was done using a inoculation loop.

Thanks goatrider you beast! Always helping and thanks & congratulations on the TC Crackatoa!

Edit- added a few more pix


Edited by Atomsplit (05/30/21 11:18 PM)


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InvisibleGoatriderS
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Re: Mold on agar? [Re: Atomsplit]
    #27329675 - 05/30/21 11:20 PM (15 days, 5 hours ago)

I really hate germinating on agar :grin:
I only shoot spores on brf pucks if i have too.
After a week transfers are done.
And the transfers from brf to agar grow pretty fast, don`t know if it`s just me.
Could be the pics, but the one has a slightly blue hue.
In a few days you`ll know.

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InvisibleAtomsplit
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Re: Mold on agar? [Re: Goatrider]
    #27329680 - 05/30/21 11:26 PM (15 days, 5 hours ago)

Do you mean a piece of fully colonized brf cake onto agar?


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InvisibleGoatriderS
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Re: Mold on agar? [Re: Atomsplit]
    #27329682 - 05/30/21 11:29 PM (15 days, 5 hours ago)

I do some little brf pucks like in the Josex`Poke.

Shoot some spores on it, it germinates fast.
Bacteria has nearly no chance on it.
First signs of growth i transfer to agar.
Works like a charm :wink:

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Re: Mold on agar? [Re: Goatrider]
    #27331730 - 06/01/21 08:09 PM (13 days, 8 hours ago)

I guess I'm being impatient this time but doesn't this growth seem off?



Time to swipe more plates?
Thank u


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Re: Mold on agar? [Re: Atomsplit]
    #27331891 - 06/01/21 10:47 PM (13 days, 6 hours ago)

That is looking like contams to me... usually when I streak a plate the mycelium looks a bit thinner and “choppy” (for lack of a better term) than that until the first transfer when it tends to get more organized and looking like cube mycelium.


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Re: Mold on agar? [Re: PitcherCrab]
    #27331925 - 06/01/21 11:32 PM (13 days, 5 hours ago)

Quote:

PitcherCrab said:
That is looking like contams to me..




Thank you! I really suck at this! :facepalm3:
I'm just flaming my loop- cooling on agar- rubbing spore then onto plate.

What seems to be a straightforward process is not as easy as I thought.

Thank you.


Edited by Atomsplit (06/01/21 11:53 PM)


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Re: Mold on agar? [Re: Atomsplit]
    #27331973 - 06/02/21 01:07 AM (13 days, 4 hours ago)

Naw man, its all good. You’ll get it! I still get contaminated plates and I’ve been doing this a while. It could just be the print you’re using is dirty. Are you working in a Still Air Box?


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“I must not fear. Fear is the mind-killer. Fear is the little death that brings total obliteration. I will face my fear. I will permit it to pass over me and through me. And when it has gone past I will turn the inner eye to see its path. Where the fear has gone there will be nothing. Only I will remain.” - Bene Gesserit Litany Against Fear


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Re: Mold on agar? [Re: PitcherCrab]
    #27332002 - 06/02/21 02:07 AM (13 days, 3 hours ago)

Quote:

PitcherCrab said:
It could just be the print you’re using is dirty. Are you working in a Still Air Box?




The print is from a shroomery vendor and I'm working in my SAB.
I was using a pretty long flimsy loop from a science supply shop. Maybe I'll make one that's less flimsy

Thanks for helping a n00b out. I appreciate it!


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Re: Mold on agar? [Re: Atomsplit]
    #27332005 - 06/02/21 02:16 AM (13 days, 2 hours ago)

Try out sterile swabs they come 2 to a package in boxes of 100 packs and they are awesome for streaking spores. I like them because I know they are 100% sterile and I don’t have to flame them before use. They’re also good if you want to make swabs later for trading, or of a variety that doesn’t drop spores like PE. Definitely worth the investment. If you keep having trouble with your print, or want to try with another one, hit me up. I’ve got a ton of extras.

Also if your not already get a little rack to work on so you’re not working on the floor of your SAB. The floor is where a lot of contams and stuff fall and collect. Same goes for working in front of a flow hood. This could help reduce nasties on your plates.

I’d also hang onto this first plate and keep an eye on it... I said I think its a contam just based on how the mycelium looks and the fact its grown that much in 6 days but I *could* be wrong. You’ll know for sure if it starts to sporulate green or if it changes color at all. Hard to tell exactly what’s what from photographs.

I hope this helps!


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“I must not fear. Fear is the mind-killer. Fear is the little death that brings total obliteration. I will face my fear. I will permit it to pass over me and through me. And when it has gone past I will turn the inner eye to see its path. Where the fear has gone there will be nothing. Only I will remain.” - Bene Gesserit Litany Against Fear


Edited by PitcherCrab (06/02/21 02:29 AM)


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InvisibleAtomsplit
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Re: Mold on agar? [Re: PitcherCrab]
    #27332084 - 06/02/21 05:02 AM (13 days, 6 minutes ago)

Quote:

I hope this helps!




Absolutely will look into buying some swabs to share prints when and if I get a grow going.

So I used bod's Tek to make a firmer inoculation loop. Worked better than my previous one. It had a lot more leverage to scrape a bit of spores onto the loop (most likely too much) and onto the agar.



Thanks for the offer on the prints pitcher! That's really really cool of you. I may take that up on you if my two prints don't work out.

Mostly thanks for the feedback on my plates. I appreciate it. There has been times where I want to quit throw in the towel but this may be one of the few chances I've ever had in my life to try a grow.

Thanks for the help!


Edited by Atomsplit (06/02/21 02:59 PM)


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Re: Mold on agar? [Re: Atomsplit]
    #27332139 - 06/02/21 06:50 AM (12 days, 22 hours ago)

I agree that growth looks contam-ish.

Mycelium is typically Rhizomorphic, or ropey.


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Re: Mold on agar? [Re: IrishGnome1] * 1
    #27332141 - 06/02/21 06:51 AM (12 days, 22 hours ago)

Quote:

IrishGnome1 said:
I agree that growth looks contam-ish.

Mycelium is typically Rhizomorphic, or ropey.




No it's typically tormentose and rhizomiphoric.


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Re: Mold on agar? [Re: DERRAYLD]
    #27332297 - 06/02/21 10:22 AM (12 days, 18 hours ago)

IME when spores first germinate the mycelia growth tend to be sort of “spotty” or patchy and kind of light. After the first of second transfer is when you can see a marked difference and it will get a lot more organized and often turn rhizomorphic.


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“I must not fear. Fear is the mind-killer. Fear is the little death that brings total obliteration. I will face my fear. I will permit it to pass over me and through me. And when it has gone past I will turn the inner eye to see its path. Where the fear has gone there will be nothing. Only I will remain.” - Bene Gesserit Litany Against Fear


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Re: Mold on agar? [Re: DERRAYLD]
    #27332393 - 06/02/21 11:39 AM (12 days, 17 hours ago)

Quote:

DERRAYLD said:
Quote:

IrishGnome1 said:
I agree that growth looks contam-ish.

Mycelium is typically Rhizomorphic, or ropey.




No it's typically tormentose and rhizomiphoric.



It's our old friend XyberSurfer21 :ban:


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Re: Mold on agar? [Re: Ashtray161]
    #27332919 - 06/02/21 07:11 PM (12 days, 9 hours ago)

Seems to soon to be able to guess


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Re: Mold on agar? [Re: PitcherCrab]
    #27332946 - 06/02/21 07:33 PM (12 days, 9 hours ago)

Quote:

PitcherCrab said:
IME when spores first germinate the mycelia growth tend to be sort of “spotty” or patchy and kind of light. After the first of second transfer is when you can see a marked difference and it will get a lot more organized and often turn rhizomorphic.



This.
I'll often take a few wedges from the germ plate just to evaluate them better on the transfer. Sometimes weak spots turn out pretty strong growth ime.


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Re: Mold on agar? [Re: LtLurker] * 1
    #27335684 - 06/04/21 10:19 PM (10 days, 6 hours ago)

Just a small update and thanks again everyone for helping. 3 days later and things looking better on the newly inoculated plates.



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Re: Mold on agar? [Re: Atomsplit]
    #27335688 - 06/04/21 10:23 PM (10 days, 6 hours ago)

Nice! This is exactly what you want to be seeing. They're germinating right along your streak line. Once they grow out a little more and it gets thicker make some transfers from the best looking spots. The T1 plates should be noticeably more vigorous.


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“I must not fear. Fear is the mind-killer. Fear is the little death that brings total obliteration. I will face my fear. I will permit it to pass over me and through me. And when it has gone past I will turn the inner eye to see its path. Where the fear has gone there will be nothing. Only I will remain.” - Bene Gesserit Litany Against Fear


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Re: Mold on agar? [Re: PitcherCrab]
    #27337538 - 06/06/21 03:03 PM (8 days, 14 hours ago)

Thanks Pitcher!

I'm looking at transferring the areas at 8 & 9 o'clock. Should I wait another day or two?

Seems like the mycelium at 9 o'clock might grow into the growth of mycelium on the 10 o'clock edge.



Thanks everyone!


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Re: Mold on agar? [Re: Atomsplit]
    #27337739 - 06/06/21 05:44 PM (8 days, 11 hours ago)

To reduce further the contamination, you could keep your inoculation loop or scalpel inside a baby jar with alcohol 90% or higher(99%) before sterilization with fire.


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Re: Mold on agar? [Re: fungusul]
    #27337839 - 06/06/21 07:08 PM (8 days, 10 hours ago)

Quote:

fungusul said:
To reduce further the contamination, you could keep your inoculation loop or scalpel inside a baby jar with alcohol 90% or higher(99%) before sterilization with fire.






Is my plate contaminated? I had the loop in iso jar. Thank you.


Edited by Atomsplit (06/06/21 07:43 PM)


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Offlineverum subsequentis
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Re: Mold on agar? [Re: Atomsplit]
    #27338212 - 06/07/21 12:48 AM (8 days, 4 hours ago)

I think you've got viable myc but can't tell yet. Looks suspicious in a few ways. I'd give it a few days.


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Re: Mold on agar? [Re: verum subsequentis]
    #27338234 - 06/07/21 01:11 AM (8 days, 3 hours ago)

Pretty fluffy stuff, but looks ok.
Probably i´d transfer from the red mark.


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Re: Mold on agar? [Re: Goatrider]
    #27338251 - 06/07/21 01:32 AM (8 days, 3 hours ago)

Thank you verum subsequentis & Goatrider! Your help is very much appreciated!

Hmm a better look at the plate at 12 o'clock


Edited by Atomsplit (06/07/21 06:09 AM)


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Re: Mold on agar? [Re: Atomsplit]
    #27341740 - 06/09/21 04:27 PM (5 days, 12 hours ago)

They say transfer off the germination plate asap.

I have a couple that is a day or two out. Obviously more growth at the spots where there is visually spores on the agar.

Do I just trust the process and start transferring from the leading edge from all the vigorous/ healthy growth spots?

All the growth seems about the same tbh. Just fuzzy/ cotton/ mold like growth - like this older plate which has already grown out.




I'm mold farming here and I hate to be negative but I'm learning this really slowly. :tongue:


Thank you


Edited by Atomsplit (06/09/21 04:41 PM)


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Re: Mold on agar? [Re: Atomsplit]
    #27341861 - 06/09/21 06:02 PM (5 days, 11 hours ago)

It's really normal to have weird fuzzy growth on the germination plate. Do transfers and see how it shakes out. All that fuzz you're worried about may very well be mycelium. :wink: BTW, what variety is this? Some, like RW, don't exhibit very much rhizomorphic mycelium.


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“I must not fear. Fear is the mind-killer. Fear is the little death that brings total obliteration. I will face my fear. I will permit it to pass over me and through me. And when it has gone past I will turn the inner eye to see its path. Where the fear has gone there will be nothing. Only I will remain.” - Bene Gesserit Litany Against Fear


Edited by PitcherCrab (06/09/21 06:03 PM)


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Re: Mold on agar? [Re: PitcherCrab]
    #27341891 - 06/09/21 06:31 PM (5 days, 10 hours ago)

Thanks again Pitcher!

Golden Teachers. I did a bunch of random transfers already - all leading edges. Was running out of plates so did 4 transfers onto a plate. :eek:

Poured 40 plates and want to let them sit for a week to see if any contamination show up.



Did a bunch of reading and thankfully found out what a "almost clear colony" looks like.

Thanks for the help! :bow2: This has not been easy.


Edited by Atomsplit (06/09/21 06:45 PM)


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Re: Mold on agar? [Re: Atomsplit]
    #27342068 - 06/09/21 08:45 PM (5 days, 8 hours ago)

For early transfers its not uncommon to do 3-4 transfers onto the same plate as you will probably just be selecting the best looking growth and transferring again to a new plate. I think you're doing great, and def doing exactly what you need to do. Just keep an eye on everything and select for the best looking mycelium! After a few transfers mycelium really starts to get more organized and rhizomorphic.

Once you're on T3 or T4 and are certain its clean take the culture to grain. :grin:


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“I must not fear. Fear is the mind-killer. Fear is the little death that brings total obliteration. I will face my fear. I will permit it to pass over me and through me. And when it has gone past I will turn the inner eye to see its path. Where the fear has gone there will be nothing. Only I will remain.” - Bene Gesserit Litany Against Fear


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Re: Mold on agar? [Re: PitcherCrab]
    #27345577 - 06/12/21 03:33 PM (2 days, 13 hours ago)

I transfered off the center piece of this plate. 3-6 o'clock area.



I really don't know if I'm selecting the best places. You guys are the only people that I can talk to this about :tongue:

Thanks!


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Re: Mold on agar? [Re: Atomsplit]
    #27345584 - 06/12/21 03:38 PM (2 days, 13 hours ago)

Can you take a photo from the top of the plate? It's hard to see what's going on...


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“I must not fear. Fear is the mind-killer. Fear is the little death that brings total obliteration. I will face my fear. I will permit it to pass over me and through me. And when it has gone past I will turn the inner eye to see its path. Where the fear has gone there will be nothing. Only I will remain.” - Bene Gesserit Litany Against Fear


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Re: Mold on agar? [Re: PitcherCrab]
    #27345593 - 06/12/21 03:47 PM (2 days, 13 hours ago)

Hey Pitcher!

From this angle I chose the 4-9 area.



Thank you again!


Edited by Atomsplit (06/12/21 03:50 PM)


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Re: Mold on agar? [Re: Atomsplit]
    #27345600 - 06/12/21 03:53 PM (2 days, 13 hours ago)

I'm liking that middle transfer. I'd take the next ones from about 3 o'clock to 7 o'clock. It's starting to look more organized. Harder to see what's going on with the others due to condensation. I'd let it grow out a tiny bit more then do another transfer. What T# is this?


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“I must not fear. Fear is the mind-killer. Fear is the little death that brings total obliteration. I will face my fear. I will permit it to pass over me and through me. And when it has gone past I will turn the inner eye to see its path. Where the fear has gone there will be nothing. Only I will remain.” - Bene Gesserit Litany Against Fear


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Re: Mold on agar? [Re: PitcherCrab]
    #27345620 - 06/12/21 04:08 PM (2 days, 12 hours ago)

This is T-1.
I went in for the transfer on the areas that were growing towards the other transfers but left the 3-5 o'clock to let it grow out.

I did 16 transfers off the T-0 and this is one of the few areas I feel ok about. 

I've been looking at these plates and reading /searching for anything to give me an idea on cleaning these up.

Thanks again Pitcher!


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OfflinePitcherCrab
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Re: Mold on agar? [Re: Atomsplit]
    #27345636 - 06/12/21 04:16 PM (2 days, 12 hours ago)

They look clean to me, I'd focus on grabbing the most organized and vigorous mycelium. :wink:


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“I must not fear. Fear is the mind-killer. Fear is the little death that brings total obliteration. I will face my fear. I will permit it to pass over me and through me. And when it has gone past I will turn the inner eye to see its path. Where the fear has gone there will be nothing. Only I will remain.” - Bene Gesserit Litany Against Fear


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Offlineverum subsequentis
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Re: Mold on agar? [Re: PitcherCrab]
    #27347429 - 06/14/21 02:54 AM (1 day, 2 hours ago)

Interesting that you think they are clean. I'm undecided as of now. I do agree that the middle one has potential and that a little growing out would make it more obvious.


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Re: Mold on agar? [Re: verum subsequentis] * 1
    #27347785 - 06/14/21 12:17 PM (16 hours, 51 minutes ago)

I chalked up the growth to looking that way because it's just a T1 and germ plates can look pretty funky. I'd still like to see some transfers and better photos of said transfers. The condensation makes it difficult to see exactly what's going on.

Atomsplit, how's it going, did you make transfers after that last post?


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“I must not fear. Fear is the mind-killer. Fear is the little death that brings total obliteration. I will face my fear. I will permit it to pass over me and through me. And when it has gone past I will turn the inner eye to see its path. Where the fear has gone there will be nothing. Only I will remain.” - Bene Gesserit Litany Against Fear


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InvisibleAtomsplit
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Re: Mold on agar? [Re: PitcherCrab]
    #27348013 - 06/14/21 04:34 PM (12 hours, 34 minutes ago)

Hey verum & pitcher!

Thanks for chiming in. All of my T-1 transfers came off of growth like this.

It's the only organized growth I see as the others look disorganized.

I've been reading tons but can find anything that looks like this. Possiblity mold?



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Re: Mold on agar? [Re: Atomsplit]
    #27348216 - 06/14/21 07:16 PM (9 hours, 52 minutes ago)

The one you took the transfers from is the best of the four. The other three are looking questionable to me in this new photo. I'd like to see how the new transfers grow out.


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“I must not fear. Fear is the mind-killer. Fear is the little death that brings total obliteration. I will face my fear. I will permit it to pass over me and through me. And when it has gone past I will turn the inner eye to see its path. Where the fear has gone there will be nothing. Only I will remain.” - Bene Gesserit Litany Against Fear


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