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OfflineAmanita virosa
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slant agar recipe
    #17211434 - 11/13/12 08:01 AM (11 years, 6 months ago)

I know low nutrient is better I have been using a diluted pda recipe.  about 1/2 strenth with a piece of popsickle stick in it.  thinking about adding a bit of sawdust to my new batch.  what is everyone else using for slant agar and do you have a favorite recipe?
I LOVE slants, btw... they are so damn pretty!!

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Offlinesolumvita
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Re: slant agar recipe [Re: Amanita virosa]
    #17222894 - 11/15/12 05:56 AM (11 years, 6 months ago)

i use the same and it seems tro be working really well!  I have thought of adding sawdust/substrate to the agar and it does work well for species certainly ABM likes it a lot.


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Re: slant agar recipe [Re: solumvita]
    #17224145 - 11/15/12 12:43 PM (11 years, 6 months ago)

I just use my normal malt recipe.
I make my slants w/ more agar than you, and get the slant all the way to the top so i dont have to shove a scalpel all the way in the tube, just the blade.


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OfflineAmanita virosa
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Re: slant agar recipe [Re: total]
    #17224599 - 11/15/12 02:20 PM (11 years, 6 months ago)

Someone gave me a few cases of these little jars.  I usually use an inoculation loop to steal just a bit of myc to sub them out....these vials are short and wide so a scalpel fits pretty well. I could see where it would be impossible to get a scalpel down those smaller test tubes though.


Edited by Amanita virosa (11/15/12 02:21 PM)

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Offlinetotal
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Re: slant agar recipe [Re: Amanita virosa]
    #17224761 - 11/15/12 03:00 PM (11 years, 6 months ago)

I use Kimble 25x150mm tubes...
And Kim-caps on top...

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OfflineOICU812
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Re: slant agar recipe [Re: total]
    #17232052 - 11/16/12 08:52 PM (11 years, 6 months ago)

I use 20x125 culture tubes & Kim Caps for my slants.

One of the long term slant recipes I use:

Long-term Preservation Medium (FDA M85)
---------------------------------------
3 g Yeast extract, 0.3%
10 g Peptone
30 g NaCl
3 g Agar
1 L Distilled water

Heat to dissolve ingredients. Dispense 4 ml portions to 13 x 100 mm screw-cap tubes. Autoclave 15 min at 121°C. Cool and tighten caps for storage. No pH adjustment is necessary.


From: FastFred's Media Cookbook


--------------------
--------------
"Democracy is two wolves and a lamb voting on what to have for lunch. Liberty is a well-armed lamb contesting the vote!" --Benjamin Franklin

"Those who give up liberty for security won't have, or deserve, either.". . . Benjamin Franklin
----> Read: The Fight of our Lives - Defeating the Ideological War Against the West - by Victor Davis Hanson

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Invisiblelipa

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Re: slant agar recipe [Re: Amanita virosa]
    #17233895 - 11/17/12 05:59 AM (11 years, 5 months ago)

19 g Agar + whatever I can find in the fridge in 1L water. :smirk: It doesn't really matter what the agar is as long as there is food. They store all the same.  With warm weather cultures I use substrate only agars for storage.

Lipa

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Re: slant agar recipe [Re: lipa]
    #17234010 - 11/17/12 07:18 AM (11 years, 5 months ago)

That's heavy on the agar, does that keep it from evaporating?

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Re: slant agar recipe [Re: falcon]
    #17234079 - 11/17/12 07:59 AM (11 years, 5 months ago)

Quote:

falcon said:
That's heavy on the agar, does that keep it from evaporating?



Thats for a liter of agar...
I use 10g/500ml

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Re: slant agar recipe [Re: lipa]
    #17234172 - 11/17/12 08:34 AM (11 years, 5 months ago)

Quote:

lipa said:
  With warm weather cultures I use substrate only agars for storage.

Lipa




Um,...what do you mean by substrate only agars??

Also, can someone explain to me how exactly you use the popsicle stick??

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Re: slant agar recipe [Re: total]
    #17234185 - 11/17/12 08:39 AM (11 years, 5 months ago)

oops, yeah. thanks

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Invisiblelipa

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Re: slant agar recipe [Re: Darkman]
    #17236319 - 11/17/12 03:52 PM (11 years, 5 months ago)

Quote:

Darkman said:
Quote:

lipa said:
  With warm weather cultures I use substrate only agars for storage.

Lipa




Um,...what do you mean by substrate only agars??

Also, can someone explain to me how exactly you use the popsicle stick??




Agars that only have straw or wood in them instead of added sugars and protein.

Lipa

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OfflineOICU812
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Re: slant agar recipe [Re: Darkman]
    #17236809 - 11/17/12 05:43 PM (11 years, 5 months ago)

Quote:

Darkman said:
Also, can someone explain to me how exactly you use the popsicle stick??




The Popsicle stick, which can also be a coffee stir stick or tongue depressor stick is simply a piece of hydrated hardwood put into the slant to help preserve the culture.

To quote RR: ". . . put a hardwood stick in each slant, and they easily last 2 to 3 years under refrigeration. If the mycelium on the agar dies, it will still revive from the wood. I use medical tongue depressors. If you don't use wood, make fresh slants every 6 to 8 months."


--------------------
--------------
"Democracy is two wolves and a lamb voting on what to have for lunch. Liberty is a well-armed lamb contesting the vote!" --Benjamin Franklin

"Those who give up liberty for security won't have, or deserve, either.". . . Benjamin Franklin
----> Read: The Fight of our Lives - Defeating the Ideological War Against the West - by Victor Davis Hanson

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OfflineAmanita virosa
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Re: slant agar recipe [Re: lipa]
    #17236917 - 11/17/12 06:15 PM (11 years, 5 months ago)

Quote:

lipa said:
19 g Agar + whatever I can find in the fridge in 1L water. :smirk: It doesn't really matter what the agar is as long as there is food.

Lipa




Great. Next we will be trying lipa's leftover pizza agar!!

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Invisiblelipa

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Re: slant agar recipe [Re: Amanita virosa]
    #17237084 - 11/17/12 06:56 PM (11 years, 5 months ago)

Stay away from that greasy of a food, but it would be nice to see if it worked.

Lipa

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Re: slant agar recipe [Re: OICU812]
    #17239321 - 11/18/12 07:42 AM (11 years, 5 months ago)

Quote:

OICU812 said:
Quote:

Darkman said:
Also, can someone explain to me how exactly you use the popsicle stick??




The Popsicle stick, which can also be a coffee stir stick or tongue depressor stick is simply a piece of hydrated hardwood put into the slant to help preserve the culture.

To quote RR: ". . . put a hardwood stick in each slant, and they easily last 2 to 3 years under refrigeration. If the mycelium on the agar dies, it will still revive from the wood. I use medical tongue depressors. If you don't use wood, make fresh slants every 6 to 8 months."




OK, thanks.  So do you put the sticks in all your slants (doesn't the stick disturb?) or only long term storage slants?

So to transfer the stick you would scrape off the agar, place in new tube and then pour fresh agar on top??

Sorry for all the questions but I plan to pour slants soon.

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OfflineDarkman
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Re: slant agar recipe [Re: lipa]
    #17239325 - 11/18/12 07:43 AM (11 years, 5 months ago)

Quote:

lipa said:
Quote:

Darkman said:
Quote:

lipa said:
  With warm weather cultures I use substrate only agars for storage.

Lipa




Um,...what do you mean by substrate only agars??

Also, can someone explain to me how exactly you use the popsicle stick??




Agars that only have straw or wood in them instead of added sugars and protein.

Lipa




OK cool, will try this.

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OfflineAmanita virosa
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Re: slant agar recipe [Re: Darkman]
    #17239352 - 11/18/12 07:59 AM (11 years, 5 months ago)

Quote:

Darkman said:
Quote:

OICU812 said:
Quote:

Darkman said:
Also, can someone explain to me how exactly you use the popsicle stick??




The Popsicle stick, which can also be a coffee stir stick or tongue depressor stick is simply a piece of hydrated hardwood put into the slant to help preserve the culture.

To quote RR: ". . . put a hardwood stick in each slant, and they easily last 2 to 3 years under refrigeration. If the mycelium on the agar dies, it will still revive from the wood. I use medical tongue depressors. If you don't use wood, make fresh slants every 6 to 8 months."




OK, thanks.  So do you put the sticks in all your slants (doesn't the stick disturb?) or only long term storage slants?

So to transfer the stick you would scrape off the agar, place in new tube and then pour fresh agar on top??

Sorry for all the questions but I plan to pour slants soon.




you put the popsicle stick into the jar prior to pouring hot agar in, then you autoclave and slant.  I dont even thing that the kind of wood matters.  it just gives the myc something to sit on.  I use "craft" sticks which i believe are fir or spruce, not hardwood and they work fine.  I have revived cultures after many months by scraping a bit of myc with a inoculation loop off the stick.  I then grow out a plate and reslant.  I usu trash the older slant or give it away since it has been opened and exposed.

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OfflineOICU812
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Re: slant agar recipe [Re: Darkman]
    #17239644 - 11/18/12 09:49 AM (11 years, 5 months ago)

Quote:

Darkman said:

OK, thanks.  So do you put the sticks in all your slants (doesn't the stick disturb?) or only long term storage slants?

So to transfer the stick you would scrape off the agar, place in new tube and then pour fresh agar on top??

Sorry for all the questions but I plan to pour slants soon.





Aloha puts a coffee stir stick in EVERY slant, regardless of media composition.  You can by a box of 1000 cheap on eBay.



Put the stick in the tube, pour the tube about 1/4 to 1/3 full of your agar mixture (the agar mixture should not be autoclaved yet), put on your caps (I use Kim-Caps) and autoclave.  When you take them out of the autoclave, while the agar is still liquid, lean the tubes against something to give the agar a SLANT to within about an inch from the top of the tube. After cooling, transfer your cultures (one culture per tube  :wink:) and parafilm the cap to the tube.  Store at 35°F - 40°F. The slanted agar makes it much easier to put the culture sample on the growing media and it also makes retrieval of the culture less of a PITA.

Aloha stores their cultures in ziplock bags that are kept in commercial refrigerators with tight temperature controls . . . and they inspect every one of them each month!  This is one of FOUR commercial refrigerators at Aloha, all of which are packed with culture slants:






Edited by OICU812 (11/18/12 09:51 AM)

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OfflineDarkman
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Re: slant agar recipe [Re: OICU812]
    #17245226 - 11/19/12 08:43 AM (11 years, 5 months ago)

Thanks for the info.

I already bought a box of 100 from a medical supply wholesaler

for the grand total of 1$ ;' )

Same supplier has small sized glass tubes with screw caps for like 25 cents!!

But I'll have to split the sticks into quarters.

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