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Offlinemattthemushroomman
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slant issues
    #24879789 - 12/30/17 09:49 PM (7 years, 18 days ago)

I have been collecting cultures for about a year and a half now and i have amassed a pretty decent collection at this point ( probably 20 species and several different strains of some). i decided i better  start making slants so i dont lose them. I bought a huge bag of 15ml pp5 centrifuge tubes to use. for some reason around 15% of them dont seem to grow at all, the transfer will get fuzzy like it should , but it never seems to expand onto the agar, i am using the same recipe as i use for my dishes. 2% MEA im using good agar from fungi perfecti. the dishes have great growth. i cannot see any kind of bacterial growth on the surface or any off color, they look exactly the same as the ones that do grow.  i made multiples of each culture and in most cases it was just 1 of three that didnt grow, so its not the cultures either.im not screwing the  caps down tight i back them off a bit and  wrap with parafilm. has anyone else experienced anything like this before? i just did a surface scrape of one that hadn't moved for 2 weeks now and  rubbed the loop on a clean agar plate to see if maybe i am just missing  a very light colored bacterial or yeast infection but i really dont think that is my problem. i just picked up 30  big 25x200 glass test tubes i am going to  make  some more masters on of all my really good cultures but i was hoping for some insight into this before i made anymore slants and waste more time and materials.  thanks in advance for your thoughts and ideas.


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InvisibleMateja
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Re: slant issues [Re: mattthemushroomman]
    #24879903 - 12/30/17 09:57 PM (7 years, 18 days ago)

So you have good growth on 2% MEA on petri dishes but then you don´t have any growth or recovery on 2% MEA in slant tubes? That sound really weird.. Did I understand correctly?


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Offlinemattthemushroomman
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Re: slant issues [Re: Mateja]
    #24879922 - 12/30/17 10:00 PM (7 years, 18 days ago)

that is correct


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InvisibleMateja
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Re: slant issues [Re: mattthemushroomman]
    #24879932 - 12/30/17 10:04 PM (7 years, 18 days ago)

That makes no sense at all. Can´t be GE issues, can't be the agar media. Can you post some pics of the problematic tubes?


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Offlinemattthemushroomman
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Re: slant issues [Re: Mateja]
    #24879938 - 12/30/17 10:05 PM (7 years, 18 days ago)

sure, let me see if i can  get  this to work on my phone, it will probly take a few mins at least.


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OfflineShu
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Re: slant issues [Re: mattthemushroomman]
    #24880018 - 12/30/17 10:12 PM (7 years, 18 days ago)

Quote:

mattthemushroomman said:
the transfer will get fuzzy like it should , but it never seems to expand onto the agar,



Most likely insufficient gas exchange imo.

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Offlinemattthemushroomman
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Re: slant issues [Re: mattthemushroomman]
    #24880028 - 12/30/17 10:16 PM (7 years, 18 days ago)

First pick is pe done 11/17  no growth. Second is H. Americanum from 12/17 and third is also H.a done same day same everything


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OfflinehamloafM
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Re: slant issues [Re: mattthemushroomman]
    #24880033 - 12/30/17 10:19 PM (7 years, 18 days ago)

The culture isn't going to last/store for nearly as long with out the popsicle stick.


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Offlinemattthemushroomman
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Re: slant issues [Re: mattthemushroomman]
    #24880039 - 12/30/17 10:21 PM (7 years, 18 days ago)

Here is what it looks like on a plate, def growing well


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Offlinemattthemushroomman
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Re: slant issues [Re: mattthemushroomman]
    #24880047 - 12/30/17 10:23 PM (7 years, 18 days ago)

Yeah, I realize now I should have a coffee sitter in there, I will with my next batch. Glad I bought 100 lol


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InvisiblePirateSwazey
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Re: slant issues [Re: mattthemushroomman]
    #24880052 - 12/30/17 10:27 PM (7 years, 18 days ago)

How did you mix your agar for the slants? I use premix and had troubles in the past where I didn't get a good mixture.

For my slants I measure normally and mix in a measuring cup, stirring with a spoon like a cup of ovaltine between filling up tubes.


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Offlinemattthemushroomman
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Re: slant issues [Re: Shu]
    #24880055 - 12/30/17 10:28 PM (7 years, 18 days ago)

i  leave the caps  partially unscrewed so they can get GE,  here is a thought on that note though,  i did these no pour, i put 7 ml in each then  filled jars with them pced and then layed in front of the hood to cool. does the pc  cook all the O2 in the slant?  maybe i just need to leave them all open fora  bit in the flow of the hood?


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Offlinemattthemushroomman
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Re: slant issues [Re: PirateSwazey]
    #24880056 - 12/30/17 10:30 PM (7 years, 18 days ago)

1 l of water, 20g malt extract and 20g of agar in a pot on the stove, once  all  well mixed i use a baster to fill each slant


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InvisibleMateja
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Re: slant issues [Re: mattthemushroomman]
    #24880063 - 12/30/17 10:33 PM (7 years, 18 days ago)

Quote:

mattthemushroomman said:
Here is what it looks like on a plate, def growing well



What culture is this?

And no the PC does not "cook" the O2, even if it did, you´d still have to let in new air when you inoculate the slants. Anyway that´s surely not the problem. GE is not a problem either so early in the transfer. This is really weird. And it doesn't seem to be unevenly mixed agar media either so this is a mystery to me :shrug:


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Offlinemattthemushroomman
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Re: slant issues [Re: Mateja]
    #24880070 - 12/30/17 10:36 PM (7 years, 18 days ago)

The plate is Hericium americanum i believe. it could possibly be coralidies. i have some growing out right now but i dont have my  air exchanges dialed in  right yet and i dont have a co2 meter to see  how bad it is in there. i need to  get a good waterproof fan i will post a pic of the fruits in a few mins


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Offlinemattthemushroomman
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Re: slant issues [Re: Mateja]
    #24880077 - 12/30/17 10:41 PM (7 years, 18 days ago)

Here is the fruits


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InvisibleMateja
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Re: slant issues [Re: mattthemushroomman]
    #24880082 - 12/30/17 10:44 PM (7 years, 18 days ago)

Quote:

mattthemushroomman said:
The plate is Hericium americanum i believe.



On the dish the culture didn't seem to grow unilaterally at all so it´s hard to imagine it would fill out the surface of the media in the tub completely. If it was cube mycelium I would give you  better answer but I don´t have experience with Hericium..


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Offlinemattthemushroomman
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Re: slant issues [Re: Mateja]
    #24880092 - 12/30/17 10:53 PM (7 years, 18 days ago)

if you lo at the second of those three pics of tubes  you can see the hericium has  spread across the surface pretty decent, the thrid pic is the same  culture but it hasnt moved. the first pic is a cube but its PE and that one is a month and a half old now. Thank you either way for your thoughts.


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OfflineShu
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Re: slant issues [Re: mattthemushroomman]
    #24880521 - 12/31/17 07:44 AM (7 years, 17 days ago)

I cannot see the type of cap you have but often they have a conical compression lip inside (vs a flat cap with a gasket) that even though you may think it's loose may still be making contact with the tube. I suggest simply giving them another quarter turn open.

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InvisiblemushboyMDiscord
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Re: slant issues [Re: Shu]
    #24880550 - 12/31/17 08:09 AM (7 years, 17 days ago)

Are slants media supposed to be less nutritious or more or doesnt matter?

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