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Offlinedrwatson
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A few Agar Questions
    #7666464 - 11/22/07 10:12 AM (16 years, 2 months ago)

I should be receiving about 50 petri dishes to get me started exploring the world of Agar work and I'm really looking forward to trying to clone a (as fresh as possible) store bought shiitake.

I realized after purchasing my petri dishes that I should have gotten the ones that have sections for individual cultures on each tray

  • Is there a simple method for making a multi-compartment petri dish with a regular petri dish so that I can have multiples on one plate?
  • Is there a basic guide I can follow step by step for isolating substrains? Or am I understanding correctly that I would basically just isolate individual growths from my Initial Dishes within their own petri dishes and then colonize a few jars of grain with these, fruit them, and then decide which one's are best based on fruit bodies, colonization times, potency, etc. . .


Thanks in advance for any help or kicks in the ass. :popcorn:


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OfflineNo_Life_G33k
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Re: A few Agar Questions [Re: drwatson]
    #7667232 - 11/22/07 02:58 PM (16 years, 2 months ago)

If you're cloning, you already habe an isolate(d strain).

No need for the divided dishes, I don know any hobbyists that use them.
They cheap by the case if you need a boat load.


--------------------
Hot Water Pasteuriztion - A How To Pictorial
http://www.shroomery.org/forums/showflat.php/Cat/0/Number/4527808/an/0/page/0

Make your own shroom chocolates
http://www.shroomery.org/forums/showflat.php/Cat/0/Number/4500682/an/0/page/5

guide with pix.


Make Your Own Magnetic Stirrer! How-To Tek w/ pix
http://www.shroomery.org/forums/showflat.php/Cat/0/Number/5865949/an/0/page/3


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OfflineRogerRabbitM
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Re: A few Agar Questions [Re: No_Life_G33k]
    #7668229 - 11/22/07 08:54 PM (16 years, 2 months ago)

You can put several specimens on each petri dish, whether or not you have them sectioned. I like the sections though. I won't ever go back to single dishes.
RR


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"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
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Offlinedrwatson
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Re: A few Agar Questions [Re: RogerRabbit]
    #7669668 - 11/23/07 10:08 AM (16 years, 2 months ago)

I was thinking about toothpicks and then saw a previous thread which answered all my questions. That thread happens to be just above this one now!

Thanks RR I'll feel confident about doing that now instead of wondering for 7 days after whether or not I fucked it up. Actually a single toothpick might just be enough to make my dishes stretch.

Are there any potential negative effects that limiting a section size might have on the Mycelium? Or perhaps maybe just certain species?


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Offlinewutang
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Re: A few Agar Questions [Re: RogerRabbit]
    #7669672 - 11/23/07 10:10 AM (16 years, 2 months ago)

dont mean to hijack your thread..but you  might get some
useful info on this...

RR, or anyone..
i have a few agar plates..and they're not rhizomorphic..at all
should i keep transfering them until they show up?

agar is awesome stuff...if you get a crappy print and inoculate you can
separate the contams and keep transfering until its gone..:thumbup:


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OfflineRogerRabbitM
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Re: A few Agar Questions [Re: wutang]
    #7669910 - 11/23/07 11:03 AM (16 years, 2 months ago)

It usually takes two or three transfers before rhizomorphs begin to show up. Below, you can see how much progress only two transfers can make.
RR



--------------------
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semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison


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Offlinewutang
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Re: A few Agar Questions [Re: RogerRabbit]
    #7670011 - 11/23/07 11:20 AM (16 years, 2 months ago)

thanks thats what i needed to know..

when i cut a slice out i noticed that there were very little
rhizomorphic growth underneath the layer of fluff
strange...


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OfflineNo_Life_G33k
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Re: A few Agar Questions [Re: wutang]
    #7678636 - 11/25/07 03:50 PM (16 years, 2 months ago)

I swear to the Gods that Roger said that he didn't use the seperated or "Y' dishes....:crazy2:


--------------------
Hot Water Pasteuriztion - A How To Pictorial
http://www.shroomery.org/forums/showflat.php/Cat/0/Number/4527808/an/0/page/0

Make your own shroom chocolates
http://www.shroomery.org/forums/showflat.php/Cat/0/Number/4500682/an/0/page/5

guide with pix.


Make Your Own Magnetic Stirrer! How-To Tek w/ pix
http://www.shroomery.org/forums/showflat.php/Cat/0/Number/5865949/an/0/page/3


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OfflineKillrX
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Re: A few Agar Questions [Re: No_Life_G33k]
    #7688042 - 11/27/07 08:20 PM (16 years, 2 months ago)

i think he said hed never go back to unseperated ones


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OfflineRogerRabbitM
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Re: A few Agar Questions [Re: KillrX]
    #7688389 - 11/27/07 09:43 PM (16 years, 2 months ago)

I prefer the three section dishes. You can run three experiments at once, saving time, money and space.
RR


--------------------
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semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison


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OfflineKillrX
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Re: A few Agar Questions [Re: RogerRabbit]
    #7688478 - 11/27/07 10:11 PM (16 years, 2 months ago)

True that just ordered 25 of em gotta try some isolations


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OfflineNo_Life_G33k
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Re: A few Agar Questions [Re: KillrX]
    #7695034 - 11/29/07 02:27 PM (16 years, 2 months ago)

Just to prove I'm OCD, not sofa king we todd ed.

rodger rabbit (Skyypilot)
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Post Number: 1319
Registered: 02-2003 Posted on Friday, December 26, 2003 - 03:28 am:

Y type? What is that? Use parafilm to seal the petri dish after inoculation.

"I feel rowdy and I don't know why. . .Excuse me, while I kiss the sky!" jimi hendrix


Kenny (Kennieyk)
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Post Number: 48
Registered: 02-2002 Posted on Friday, December 26, 2003 - 03:38 am:

Its a petri dish that is sectioned of in 3 parts . Like a Y shape ! The y section is only in the bottom half . I guess I could just use the top . Where can I get the parafilm?


rodger rabbit (Skyypilot)
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Registered: 02-2003 Posted on Friday, December 26, 2003 - 04:14 am:

You can get proper petri dishes and parafilm here: http://www.fungi.com/tools/containers.html
"I feel rowdy and I don't know why. . .Excuse me, while I kiss the sky!" jimi hendrix

<!--end quote>

it was on mycotopia, and you had the pic of the fantastic invito pins


--------------------
Hot Water Pasteuriztion - A How To Pictorial
http://www.shroomery.org/forums/showflat.php/Cat/0/Number/4527808/an/0/page/0

Make your own shroom chocolates
http://www.shroomery.org/forums/showflat.php/Cat/0/Number/4500682/an/0/page/5

guide with pix.


Make Your Own Magnetic Stirrer! How-To Tek w/ pix
http://www.shroomery.org/forums/showflat.php/Cat/0/Number/5865949/an/0/page/3


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OfflineRogerRabbitM
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Re: A few Agar Questions [Re: No_Life_G33k]
    #7696465 - 11/29/07 07:13 PM (16 years, 2 months ago)

Quote:

Y type? What is that?




Wow, you did do some searching. I had never heard of the three section dishes being called "Y type", so I didn't catch on at first to what the guy was saying.

I keep some of both types around, but usually use the three section ones.
RR


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Download Let's Grow Mushrooms



semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
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Offlinedrwatson
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HELP! A few Agar Questions [Re: RogerRabbit]
    #7698848 - 11/30/07 12:59 PM (16 years, 2 months ago)

Okay, I'm a dumbass.  Truly NooB.

Well i can't find AgarAgar ANYWHERE in my city so i decided i would try the "pink one" Fruit Pectin instead of Agar  :nonono:

Well now I know that it only stays solid until the Mycelium start to eat it and then it turns to liquid and just sloshes around the petri dish if/when moved.  :nonono:

How can I save it?
I have a dish with a Mazapatec culture started and the whole thing is swimming in PDY solution :sad:

Can I just spoon it into a WBS jar?  Would it be better let it colonize the entire petri first?  If so, would it stop swimming around at some point when it's consumed most of the nutrient solution?

edit:Any suggestions are much appreciated as I saved this little guy from a syringe I thought to be bunk or contaminated, and of 12 cultures started from said syringe only this one grew.  So i'd like to save it.


Edited by drwatson (11/30/07 01:03 PM)


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Re: HELP! A few Agar Questions [Re: drwatson]
    #7699220 - 11/30/07 02:06 PM (16 years, 2 months ago)

I would pour that mess into a LC of karo and water sterilized of course
good luck


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Offlinedrwatson
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Re: HELP! A few Agar Questions [Re: orchidfanatic]
    #7699256 - 11/30/07 02:13 PM (16 years, 2 months ago)

That isn't a bad idea . . . it would be a bit risky for contams. but then again so will anything else i do to save it. That might actually be the quickest method with the most "fluid" action. (no pun intended)


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Re: HELP! A few Agar Questions [Re: drwatson]
    #7699320 - 11/30/07 02:30 PM (16 years, 2 months ago)

or you can let it sit and get some vendor to send you either prepoured plates or malt extract agar mix and make some more plates transfer some mycelium onto a real agar plate. do a search for agar substitutes someone on here made some kind of flour agar that seems to work
good luck


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Re: HELP! A few Agar Questions [Re: orchidfanatic]
    #7699325 - 11/30/07 02:31 PM (16 years, 2 months ago)

I think mushymart carries prepoured agar plates


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Re: HELP! A few Agar Questions [Re: drwatson]
    #7699482 - 11/30/07 03:15 PM (16 years, 2 months ago)

Quote:

drwatson said:
Well i can't find AgarAgar ANYWHERE in my city so i decided i would try the "pink one" Fruit Pectin instead of Agar  :nonono:

Well now I know that it only stays solid until the Mycelium start to eat it and then it turns to liquid and just sloshes around the petri dish if/when moved.  :nonono:




Your experience is exactly why agar substitutes don't work.  All the discussion of substitutes is mostly a waste of time.  Even the best substitutes, like guar gum, have problems that make them not worth the time or risk to your experiments.  Any money you save is going to be more than made up for in wasted time, frustration, and failed experiments.

Agar is not hard to find.  If you can't find it locally you probably aren't looking hard enough.  It's also easy enough to simply mail order it.  There was a recent thread on sources for cheap low/food grade agar.  That should work well and ordering online is going to save you quite a bit of money over any source I've been able to find.

As far as saving your cultures they are basically LCs now, so just treat them as such.  Transfer a few small bits to another LC, or your other option is to get some agar and plate out a mL or two onto a new plate.  If you want to isolate anything then use some sterilized toothpicks or other tool to transfer the myc to new dishes.


-FF


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Re: HELP! A few Agar Questions [Re: fastfred]
    #7699522 - 11/30/07 03:26 PM (16 years, 2 months ago)

i would rather find some agar locally. I checked the phone book and there are some Asian markets hiding around town that I had no idea existed so I'm hitting them up this weekend.

I would prefer to transfer to another plate w/real PDYA although I'll probably just transfer to a LC jar instead. Or do half and half if I find Agar this weekend.

I'll never try anything other than agar again that's for sure.

edit: By the way, thanks for the replies guys I really appreciate the help and suggestions.


Edited by drwatson (11/30/07 03:27 PM)


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Offlinedrwatson
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Re: HELP! A few Agar Questions [Re: fastfred]
    #7699539 - 11/30/07 03:31 PM (16 years, 2 months ago)

Quote:

fastfred said:


If you want to isolate anything then use some sterilized toothpicks or other tool to transfer the myc to new dishes.


-FF




This is very helpful thank you FF! As obvious as it seems I don't think I would have thought to use a toothpick initially.


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OfflineRogerRabbitM
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Re: HELP! A few Agar Questions [Re: drwatson]
    #7699597 - 11/30/07 03:44 PM (16 years, 2 months ago)

When you go to an oriental grocery, ask for 'auger-auger' as that's how they pronounce it.
RR



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Re: HELP! A few Agar Questions *DELETED* [Re: RogerRabbit]
    #7700355 - 11/30/07 06:44 PM (16 years, 2 months ago)

Post deleted by Chemy

Reason for deletion: If you are sure you want to do this, click the button below. Yes, I want to delete this post.



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Re: HELP! A few Agar Questions [Re: Chemy]
    #7701481 - 12/01/07 12:12 AM (16 years, 2 months ago)



this is what it looks like liquefied.  not sure if you can tell by this picture, as my camera sucks ass. 

An interesting side note is that I think I see the head of a gorilla in the lower left and the mycelium looks JUST LIKE a vagina. 
I wish my camera could capture it.  hahaha :thumbdown:


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Re: HELP! A few Agar Questions [Re: drwatson]
    #7702095 - 12/01/07 08:52 AM (16 years, 2 months ago)

I see the gorilla ! too bad its not a shape of jesus or the virgin mary !LOL


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Re: HELP! A few Agar Questions [Re: orchidfanatic]
    #7703675 - 12/01/07 05:44 PM (16 years, 2 months ago)

> As obvious as it seems I don't think I would have thought to use a toothpick initially.

It's a great trick. Instead of trying to sterilize loops or pics you can just sterilize a box of toothpicks with some foil over them and you have a bunch of sterile, disposable picks to use.


-FF


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