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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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agar hot pour
    #13023758 -

Hello,

I pour hot agar over some petri dishes that have a bacterial
contamination.  I hope to take a clean transfer.

I will keep an eye on these dishes, but wanted to ask
how long it usually takes for the recovering mycelia to
break back through to the new surface?

Thanks!

JD


--------------------
Boyd Rice told my brother that life is a corny pack of freesakes

Myco-tek.org

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Re: agar hot pour [Re: Javadog]
    #13023772 -

You poured hot agar onto an already inoculated dish?  You think its going to grow thru the agar?  myc grows 2d on the top of agar I dont think it will do anything but kill your myc.  Why didnt you just cut a transfer and transfer it?  I think im lost as to wat you want to do:shrug:


--------------------
Ask  AMU    for    the  best    mycology    advice  out  there
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I don't mean to boast, but dam if I don't brag.

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Re: agar hot pour [Re: truskool]
    #13023797 -

It works. I've done it to clean cultures, but not with bacteria contams. So, I can't say for sure what will break through to the surface first. But, when I did this with a particularly dirty culture I was trying to clean the myc broke through to the surface about 12 hours before the trich.
Tao


--------------------
Follow the light
The Light is your guide.

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Re: agar hot pour [Re: fungus_tao]
    #13023808 -

fungus_tao said:
It works. I've done it to clean cultures, but not with bacteria contams. So, I can't say for sure what will break through to the surface first. But, when I did this with a particularly dirty culture I was trying to clean the myc broke through to the surface about 12 hours before the trich.
Tao


Interesting I may try it.  But why dont you just transfer from the dirty dish to a clean one?  as long as there is some visible myc your good and if there isnt how do you think that the myc will break thru the contam?


--------------------
Ask  AMU    for    the  best    mycology    advice  out  there
  Roll it :burnone:While I troll it.  :maximumtrolling:

I don't mean to boast, but dam if I don't brag.

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Re: agar hot pour [Re: Javadog]
    #13023832 -

I lol'd at your sig.

In my experience bacteria grows fairly slowly and it somewhat easy to battle just be REALLY careful when handling a bacterial culture on petri dishes. You could get really really really really sick.

I suggest if you are out of petri dishes(i'm assuming you are which is why you don't just transfer a section of mycelium) that you go and pour some new agar into a quart jar or something that can be sterilized.

Don't fuck around with contams on dishes man. I can't stress enough how dangerous that is.

If you have more clean dishes, then pour the agar into them and sterilize either a scalpel or a piece of wire made into an inoculation loop to transfer a piece of mycelium. Even if the transfer carries some of the contaminate you will be a lot better off singling it out slowly rather than taking risks.


--------------------
By the livin' Gawd that made you,
    You're a better man than I am, Gunga Din

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Re: agar hot pour [Re: truskool]
    #13023837 -

truskool said:
fungus_tao said:
It works. I've done it to clean cultures, but not with bacteria contams. So, I can't say for sure what will break through to the surface first. But, when I did this with a particularly dirty culture I was trying to clean the myc broke through to the surface about 12 hours before the trich.
Tao


Interesting I may try it.  But why dont you just transfer from the dirty dish to a clean one?  as long as there is some visible myc your good and if there isnt how do you think that the myc will break thru the contam?


It was just one of the things that one can try.

When taking transfers it can be hard to not also get contams.

This is my first time though...I had to choose one path
of the other.

Tao: That 12 hour window is not large.

I need to know how long before this window opened.

How long after the hot-pour did your mycelium break through?

(I probably put a few mms of agar in the 2nd pour)

Thanks!


--------------------
Boyd Rice told my brother that life is a corny pack of freesakes

Myco-tek.org

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Re: agar hot pour [Re: truskool] * 1
    #13023851 -

It's a method of cleaning called 'chasing'. By pouring fresh agar on top of a contaminated culture you are creating something that they must grow through, not just across. This is where mycellium often has the upper hand. It depends on the contam your chasing of course, but if your mycellium wins the race you transfer to a new dish for the win. There are a few reasons to do this vs transferring. If the culture is extremely contaminated it's pretty much your only option left. Also, it doesn't require the dish being open for more than a few seconds to pour hot agar and every second counts when you've got exposed mold spores in front of you.

Edit-
Quote:
Tao: That 12 hour window is not large.


No, it's not very large but it's still an opportunity. The window gets bigger the more agar you pour on top. If you poured a very thin layer then the window is even less. The best way to do this is in a culture tube, or even  a jar so you can increase the distance of the chase to make it more effective.
Tao


--------------------
Follow the light
The Light is your guide.

Edited by fungus_tao (08/08/10 11:28 PM)

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Re: agar hot pour [Re: Javadog] * 2
    #13023921 -

12 to 36 hours, depending on how thick the second layer is.  When it breaks through, take a scalpel and 'shave' the mycelium off, taking none of the agar.  You may need to repeat when you get it transferred if the bacteria shows up.  If not, you're good to go.

For those who don't know WTF he's talking about, posted about 5 years ago.
RR


--------------------
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."
Thomas Edison

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Re: agar hot pour [Re: RogerRabbit]
    #13024071 -

hmm....how hot can this agar be? Obviously you have to hold the bottle so it can't be that hot, and if it works it's not killing the mycelium so....

Interesting tip!


--------------------
and let it be so Praise Psilocybe

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Re: agar hot pour [Re: rustycobwebs] * 1
    #13024151 -

Well, agar sets at 95F. I'm not sure exactly what temp it's at when I pour, but you don't want it hot to the touch or you get too much condensation. At any rate you wouldn't be able to pick up the flask to pour if the agar solution were hot enough to kill mycelium.
Tao


--------------------
Follow the light
The Light is your guide.

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Re: agar hot pour [Re: rustycobwebs]
    #13024180 -

rustycobwebs said:
hmm....how hot can this agar be? Obviously you have to hold the bottle so it can't be that hot


Pretty darned hot.  I use a paper towel to protect my latex gloved hands from getting burned when I do this.  I'd estimate at around 140F/60C.  It cools down pretty fast, and the mushroom mycelium survives, but it 'pasteurizes' the bacteria, knocking it back just long enough for the mushroom mycelium to recover.  Be sure to be right there when it pops through the top though, because the bacteria isn't far behind.  If there's a LOT of bacteria, use antibiotic agar.  Bear in mind, there has to be at least some mushroom mycelium for this to work.
RR


--------------------
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."
Thomas Edison

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Re: agar hot pour [Re: RogerRabbit]
    #13024253 -

RogerRabbit said:
12 to 36 hours, depending on how thick the second layer is.  When it breaks through, take a scalpel and 'shave' the mycelium off, taking none of the agar.  You may need to repeat when you get it transferred if the bacteria shows up.  If not, you're good to go.

For those who don't know WTF he's talking about, posted about 5 years ago.
RR



:hammertime:


--------------------
     
How I Get Stuff done. - My Reference Guide. - My Grows.

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Re: agar hot pour [Re: RogerRabbit]
    #13024355 -

RogerRabbit said:
12 to 36 hours, depending on how thick the second layer is.  When it breaks through, take a scalpel and 'shave' the mycelium off, taking none of the agar.  You may need to repeat when you get it transferred if the bacteria shows up.  If not, you're good to go.

For those who don't know WTF he's talking about, posted about 5 years ago.
RR


Yeah...I posted about, hoping to lure you out with that pointer.  ;0)

(at least it was old enough for my difficulty in finding
is, again!, was understandable)

I poured at about 140 and will take a good look at the
plates in front of my flowhood tomorrow morning.

Thanks!

JD


--------------------
Boyd Rice told my brother that life is a corny pack of freesakes

Myco-tek.org

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Re: agar hot pour [Re: Javadog]
    #13039631 -

Hello all,

I wanted to let you know that yesterday I check the dishes
and the Cambodian and the African Transeki has grown through
to the top of the agar.  It was cool, the growth reaches
the top making something of an impression of the growth
below on the surface.  It is easier to see from an angle.

The Elephant Dung may have a more aggressive strain of
bacteria in it (maybe some dung got into the syringe ;0)
because the pattern of contamination on the lower level
seemed to leap to the new surface.

I made transfer of the two good dishes to two new dishes each.
I also started a new dish of ED, in the hopes that I might do better...

We will see if I get clean growth...I do not mind transferring
again, but I hope to be able to do it without another hot pour,
as I already have dishes ready.

Take care,

JD


--------------------
Boyd Rice told my brother that life is a corny pack of freesakes

Myco-tek.org

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Re: agar hot pour [Re: Javadog]
    #13048995 -

BTW: taking that first "impression" required taking some agar
and so I went back last night and used my loop to shave off
just some mycelia. (it had grown through to the point that
a sample was easy)

I feel pretty good about both passes.  The first transfers are
growing nicely, and the second was very clean.

My main concern is that the plates were not contaminated
during the process, but that is why I built my baby:


--------------------
Boyd Rice told my brother that life is a corny pack of freesakes

Myco-tek.org

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Re: agar hot pour [Re: Javadog]
    #13144741 -

Hello all,

I wanted to let you know that the hot pour worked for both strains. 

This is a photo of the African Transeki transfer:


Take care,

JD


--------------------
Boyd Rice told my brother that life is a corny pack of freesakes

Myco-tek.org

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Re: agar hot pour [Re: Javadog]
    #13145924 -

:highfive:


--------------------
     
How I Get Stuff done. - My Reference Guide. - My Grows.

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Re: agar hot pour [Re: hamloaf]
    #13145941 -

:confused2::bow2::cheer::strokebeard::boobs::bow:

that is really cool


--------------------
The above post is fictional, hypothetical, or downright nonsensical.


:moon:  :moon:  :moon:  :moon:    :moon:

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Re: agar hot pour [Re: Javadog]
    #13145953 -

Javadog said:
This is a photo of the African Transeki transfer:





Gorgeous!

You know the file name says 'Cambodian', right?


--------------------
You make it all possible. Doesn't it feel good?

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Re: agar hot pour [Re: Doc_T]
    #13146101 -

Wow....I just got up...that lost me.

just a moment.....Checking the notes.

19 == ED
20 == Cambodian
21 == African Transeki
22 == Jalisco

OK, so I got them switched!  LOL.  They both cleaned up nicely.  The *Cambodian* was the nicer picture though.

Good Catch!

JD


--------------------
Boyd Rice told my brother that life is a corny pack of freesakes

Myco-tek.org

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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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