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Myyco.com Shop: Isolated Cubensis Liquid Culture For Sale

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OfflineGod Stamet
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Registered: 07/01/12
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slants didn't solidify.
    #17561339 - 01/17/13 08:41 AM (11 years, 4 months ago)

at least most of them didn't. i followed the directions for the mix,  stirred it vigorously. woke up today and some(most) didn't solidify.i'm using 25 by 150 mm standard grade culture slants with PDA. any thoughts or advice would e appreciated. also when atoclaved i put the slants inside ball pint jars then into the pc for 20 min at 15 PSI.


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I have caught the Mycology bug! Not just here for the ride, i'm here for life.

How I do my bulk substrate

just another mushroom capsule TEK

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OfflineJeff
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Re: slants didn't solidify. [Re: God Stamet]
    #17561639 - 01/17/13 09:59 AM (11 years, 4 months ago)

Did you dissolve (boil) the agar before you put it in your culture tubes?
I would suspect that it seperated out of solution before you got it fully transfered.  Well that is if you didn't boil it first.


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Myco-tek

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OfflineGod Stamet
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Re: slants didn't solidify. [Re: Jeff]
    #17561766 - 01/17/13 10:31 AM (11 years, 4 months ago)

I mixed the solution then poured into the slants then pc'd the slants at 15 psi for 20 minutes. let them cool then set at a slant. 4 out of 12 did solidify. So i should boil the mix,pour, then pc? when i do petri dishes i just mix in a bottle pc for 35 minutes then pour to dishes with no problems. first time with slants though. any input is appreciated cause agar ain't cheap lol. i'm gonna pc them again for 30 minutes at 15psi and wait longer for them to cool. hopefully that does it. any info from you guys in the mean time would be great.

thanks--GS


--------------------
I have caught the Mycology bug! Not just here for the ride, i'm here for life.

How I do my bulk substrate

just another mushroom capsule TEK

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OfflineGod Stamet
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Re: slants didn't solidify. [Re: God Stamet]
    #17561817 - 01/17/13 10:45 AM (11 years, 4 months ago)

round 2! here's what i did this time. i put the slants in pint size ball jars like last time only this time i put water in the ball jars to. last time i just put the slants in an empty jar then into the water bath inside the pc. also this time i'm going to give them 30 minutes instead of 20 and wait at least a couple hours to take out and put at a slant. last time i waited less then an hour to pull them out. hopefully that does it! like i said before, any input from experienced slant users would be great


--------------------
I have caught the Mycology bug! Not just here for the ride, i'm here for life.

How I do my bulk substrate

just another mushroom capsule TEK

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OfflineJeff
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Re: slants didn't solidify. [Re: God Stamet]
    #17561820 - 01/17/13 10:46 AM (11 years, 4 months ago)

Quote:

God Stamet said:
So i should boil the mix,pour, then pc?



That is what I do.  If it isn't boiled first or continuously mixed as you pour the tubes, the agar will sink to the bottom of the mixing container and some of your slants will only contain water or a small amount of agar.

I don't boil when doing plates as the mixture becomes homogeneous as it is being pc'd due to the mix all being in one container.


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Myco-tek

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OfflineGod Stamet
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Re: slants didn't solidify. [Re: Jeff]
    #17561928 - 01/17/13 11:17 AM (11 years, 4 months ago)

interesting. i was just following RR's video. i will try that with my next batch for sure. and it did look like the agar was on the bottom of the slants that didn't solidify. do you think pc'ing them again is gonna help? either way i'm in the middle of doing it so i'll report my results. anyone else?

thanks jeff! GS


--------------------
I have caught the Mycology bug! Not just here for the ride, i'm here for life.

How I do my bulk substrate

just another mushroom capsule TEK

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OfflineOrtho
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Re: slants didn't solidify. [Re: God Stamet]
    #17563147 - 01/17/13 03:34 PM (11 years, 4 months ago)

Yeah, now i put it in the blender before i pc it. I also shake it before each plate.

Edited by Ortho (01/17/13 03:35 PM)

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OfflineGod Stamet
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Re: slants didn't solidify. [Re: Ortho]
    #17566810 - 01/18/13 08:51 AM (11 years, 3 months ago)

thanks again jeff, i made a new batch. i pressure cooked the whole batch in a quart jar for 20 minutes first. then i swirled the mix well then poured into my slants. they all solidified perfect. now i'm just gonna re-autoclave for 20 minutes and slant them. great advice, saved me some time and money by pointing out the problem for sure.

cheers! GS


--------------------
I have caught the Mycology bug! Not just here for the ride, i'm here for life.

How I do my bulk substrate

just another mushroom capsule TEK

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OfflineTerry M
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Re: slants didn't solidify. [Re: God Stamet]
    #17566827 - 01/18/13 08:56 AM (11 years, 3 months ago)

I don't cook my slant agar before use. But mixing is really important. Otherwise the agar can stratify in the bottle, and you'll get some slants more solid than others. And adding a bit of extra agar won't hurt, since brands of agar seem to vary somewhat in gelling capacity.

Always mix the water into the agar, first using small amounts of water while stirring well, and then more water. I mix mine with a spoon, in a 4-cup Pyrex measuring cup. Chefinainteasy has to know all about this. It's the technique chefs use to make perfectly thickened, lump-free sauces!


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Liberté, égalité, humidité.

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OfflineGod Stamet
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Re: slants didn't solidify. [Re: Terry M]
    #17567150 - 01/18/13 10:27 AM (11 years, 3 months ago)

thanks terry. when i mix my agar i do follow all those rules. i learned fromm RR's videos like most of us here. i guess because i give the bottle a good swirl before pouring plates i've avoided this in the past. first time with slants, so i'm not to upset about making a mistake. now i know! thanks again to everyone for helping me solve my problem.


--------------------
I have caught the Mycology bug! Not just here for the ride, i'm here for life.

How I do my bulk substrate

just another mushroom capsule TEK

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Invisible36fuckin5
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Registered: 08/11/03
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Loc: Diving into Mystical Territori...
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Re: slants didn't solidify. [Re: God Stamet]
    #17568098 - 01/18/13 01:54 PM (11 years, 3 months ago)

RR fails to mention in his video that not all agar will dissolve at room temp. Most of it needs heated to fully dissolve.

I make it a habit of boiling my water, stirring in my ingredients then you can either jar it up or pour it into slants and PC.


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Pat The Bunny said:
A punk rock song won't ever change the world, but I can tell you about a couple that changed me.

bodhisatta said:
i recommend common sense and figuring it out.

These are the TEKs I use. They're all as cheap and easy as possible, just like your mom.

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OfflineGod Stamet
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Re: slants didn't solidify. [Re: 36fuckin5]
    #17568208 - 01/18/13 02:15 PM (11 years, 3 months ago)

yup, and that's what i'll be doing from now on with my PDA. maybe MEA dissolves easier??? either way lesson learned. surprised i couldn't find any post's like this.


--------------------
I have caught the Mycology bug! Not just here for the ride, i'm here for life.

How I do my bulk substrate

just another mushroom capsule TEK

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OfflineJeff
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Re: slants didn't solidify. [Re: God Stamet]
    #17568237 - 01/18/13 02:23 PM (11 years, 3 months ago)

Actually no agar dissolves at room temperature that I am aware of.  Tic gums offers one that will disolve at 180 f.  Every other is right at boiling.


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Myco-tek

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Invisible36fuckin5
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Posts: 12,091
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Re: slants didn't solidify. [Re: God Stamet]
    #17568704 - 01/18/13 03:52 PM (11 years, 3 months ago)

Quote:

God Stamet said:
yup, and that's what i'll be doing from now on with my PDA. maybe MEA dissolves easier???




Nope. It's not really the nutrient that's the problem. It's the agar agar itself. And it all needs heated.


--------------------
Pat The Bunny said:
A punk rock song won't ever change the world, but I can tell you about a couple that changed me.

bodhisatta said:
i recommend common sense and figuring it out.

These are the TEKs I use. They're all as cheap and easy as possible, just like your mom.

Extras: Filter Print Post Top
OfflineGod Stamet
wannabe Stamets


Registered: 07/01/12
Posts: 774
Last seen: 9 years, 6 months
Re: slants didn't solidify. [Re: 36fuckin5]
    #17570385 - 01/18/13 09:34 PM (11 years, 3 months ago)

i know we've been in this age for a while now....but information through experience is a beautiful thing. thanks for sharing i really appreciate it


--------------------
I have caught the Mycology bug! Not just here for the ride, i'm here for life.

How I do my bulk substrate

just another mushroom capsule TEK

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Offlinetotal
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Re: slants didn't solidify. [Re: God Stamet]
    #17627994 - 01/29/13 09:34 AM (11 years, 3 months ago)

An immersion blender and a boiling pot of water makes light work of the agar...:smirk:

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Myyco.com Shop: Isolated Cubensis Liquid Culture For Sale


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