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OfflineSma11mau5
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Registered: 08/15/12
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Last seen: 10 years, 7 months
Agar transfers showing super slow activity, sometimes drying out
    #17314913 - 12/01/12 01:50 PM (12 years, 1 month ago)

Recently interested in trying to learn agar work, cloning etc.

Just finished first sleeve of plates, and had no real success, each time a plate was used once the specimen was on the plate(pins, center stem tissue, spores, liquid culture) sometimes some activity would be witnessed but usually by the time any mycellial growth worth transferring could be seen the plate seemed to be drying out as the agar would peel away from the sides of the plate and shrink and kind of turn yellow ish, not contam for sure, but seems like its drying out...

If anyone can offer help with this would really appreciate it

Maybe are the plates poured to thin/shallow??

As far as the agar it is a premixed that came from my local hobby science store, it doesn't specify if it's PDA or MEA or whatever just says premixed agar.

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OfflinePussyFart
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Re: Agar transfers showing super slow activity, sometimes drying out [Re: Sma11mau5]
    #17314921 - 12/01/12 01:52 PM (12 years, 1 month ago)

How are you storing the dishes?

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Offlineskippydude
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Re: Agar transfers showing super slow activity, sometimes drying out [Re: Sma11mau5]
    #17314931 - 12/01/12 01:56 PM (12 years, 1 month ago)

PICTURES:waits:

No ingredients on the bottle?

May be pure agar with no nutes.

How much do you use for 1000ml batch?

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OfflineTmethylM
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Re: Agar transfers showing super slow activity, sometimes drying out [Re: skippydude]
    #17314960 - 12/01/12 02:04 PM (12 years, 1 month ago)

Tape them up after transfer. It will stop dying out and mycelium will be able to colonize.
Pouring the agar a little thicker will help.


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OfflineSma11mau5
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Registered: 08/15/12
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Re: Agar transfers showing super slow activity, sometimes drying out [Re: Tmethyl]
    #17320031 - 12/02/12 11:12 AM (12 years, 1 month ago)

Use the poor mans glad sliced into ghetto paradigm rolls and was wrapping tightly around the dish seals but thought that would suffocate them, so I started doing a few straps around like a ribbon on a xmas present.

As far as the agar, it doesn't come with ingredients because the sci shop it came from packs it to order, they give you a little old school film canister of it with a little instruction sheet, neither says what type of agar it is.. Was so excited they had ready to use agar that the question was forgotten when in the shop.

I used a half recipe because the shop said you could do 40 plates with the whole batch of agar... So half the agar with 500 ml water, pressure cooked, poured into plates in the glovebox - so maybe in the future the whole recipe should be used for a 20 plate sleeve?

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OfflineMontock
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Re: Agar transfers showing super slow activity, sometimes drying out [Re: Sma11mau5]
    #17320041 - 12/02/12 11:15 AM (12 years, 1 month ago)

Quote:

Sma11mau5 said:
Use the poor mans glad sliced into ghetto paradigm rolls and was wrapping tightly around the dish seals but thought that would suffocate them, so I started doing a few straps around like a ribbon on a xmas present.

As far as the agar, it doesn't come with ingredients because the sci shop it came from packs it to order, they give you a little old school film canister of it with a little instruction sheet, neither says what type of agar it is.. Was so excited they had ready to use agar that the question was forgotten when in the shop.

I used a half recipe because the shop said you could do 40 plates with the whole batch of agar... So half the agar with 500 ml water, pressure cooked, poured into plates in the glovebox - so maybe in the future the whole recipe should be used for a 20 plate sleeve?



take a sheet of that glad, hold it to your mouth and try to breathe threw it, this same effect happens with your dishes

if your not providing filterd ge with parafilm you might as well use scotch tape


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OfflineSma11mau5
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Registered: 08/15/12
Posts: 98
Last seen: 10 years, 7 months
Re: Agar transfers showing super slow activity, sometimes drying out [Re: Montock]
    #17320075 - 12/02/12 11:22 AM (12 years, 1 month ago)

Quote:

Montock said:
Quote:

Sma11mau5 said:
Use the poor mans glad sliced into ghetto paradigm rolls and was wrapping tightly around the dish seals but thought that would suffocate them, so I started doing a few straps around like a ribbon on a xmas present.

As far as the agar, it doesn't come with ingredients because the sci shop it came from packs it to order, they give you a little old school film canister of it with a little instruction sheet, neither says what type of agar it is.. Was so excited they had ready to use agar that the question was forgotten when in the shop.

I used a half recipe because the shop said you could do 40 plates with the whole batch of agar... So half the agar with 500 ml water, pressure cooked, poured into plates in the glovebox - so maybe in the future the whole recipe should be used for a 20 plate sleeve?



take a sheet of that glad, hold it to your mouth and try to breathe threw it, this same effect happens with your dishes

if your not providing filterd ge with parafilm you might as well use scotch tape




Glad cling wrap is supposed to be great for plates...according to RR and others:

http://www.shroomery.org/forums/showflat.php/Number/5646666

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Offlineskippydude
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Re: Agar transfers showing super slow activity, sometimes drying out [Re: Sma11mau5]
    #17320145 - 12/02/12 11:35 AM (12 years, 1 month ago)

Quote:

Sma11mau5 said:
Use the poor mans glad sliced into ghetto paradigm rolls and was wrapping tightly around the dish seals but thought that would suffocate them, so I started doing a few straps around like a ribbon on a xmas present.

As far as the agar, it doesn't come with ingredients because the sci shop it came from packs it to order, they give you a little old school film canister of it with a little instruction sheet, neither says what type of agar it is.. Was so excited they had ready to use
I used a half recipe because the shop said you could do 40 plates with the whole batch of agar... So half the agar with 500 ml water, pressure cooked, poured into plates in the glovebox - so maybe in the future the whole recipe should be used for a 20 plate sleeve?



Quote:

Sma11mau5 said:
Use the poor mans glad sliced into ghetto paradigm rolls and was wrapping tightly around the dish seals but thought that would suffocate them, so I started doing a few straps around like a ribbon on a xmas present.

As far as the agar, it doesn't come with ingredients because the sci shop it came from packs it to order, they give you a little old school film canister of it with a little instruction sheet, neither says what type of agar it is.. Was so excited they had ready to use agar that the question was forgotten when in the shop.

I used a half recipe because the shop said you could do 40 plates with the whole batch of agar... So half the agar with 500 ml water, pressure cooked, poured into plates in the glovebox - so maybe in the future the whole recipe should be used for a 20 plate sleeve?




500ml should do 25 no problem, 20 for sure.

If you fill more than a third of the dish, you  will have problems with transfers not having enough head space.

Sounds like you have plain agar no nutrients.

How many grams is used per 1000ml?

If it is around 20 grams it is likely plain agar.

If it is more like 40 grams it is probably MEA.

I have been using saran wrap, the only problem I have had is with a couple dishes I refrigerated right side up, the collected a tablespoon of water in them.

http://www.shroomery.org/forums/showflat.php/Number/17200242/page/2

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OfflineMontock
Time = $
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Re: Agar transfers showing super slow activity, sometimes drying out [Re: Sma11mau5]
    #17321442 - 12/02/12 03:27 PM (12 years, 1 month ago)

Quote:

Sma11mau5 said:
Quote:

Montock said:
Quote:

Sma11mau5 said:
Use the poor mans glad sliced into ghetto paradigm rolls and was wrapping tightly around the dish seals but thought that would suffocate them, so I started doing a few straps around like a ribbon on a xmas present.

As far as the agar, it doesn't come with ingredients because the sci shop it came from packs it to order, they give you a little old school film canister of it with a little instruction sheet, neither says what type of agar it is.. Was so excited they had ready to use agar that the question was forgotten when in the shop.

I used a half recipe because the shop said you could do 40 plates with the whole batch of agar... So half the agar with 500 ml water, pressure cooked, poured into plates in the glovebox - so maybe in the future the whole recipe should be used for a 20 plate sleeve?



take a sheet of that glad, hold it to your mouth and try to breathe threw it, this same effect happens with your dishes

if your not providing filterd ge with parafilm you might as well use scotch tape




Glad cling wrap is supposed to be great for plates...according to RR and others:

http://www.shroomery.org/forums/showflat.php/Number/5646666



thats 6 years old


its not


try my breathing test or ask rr nowadays


--------------------
Looking for Wife, please pm me your picture and list of skillz.


I'm wrong all the time, my posts are subject to being wrong at any  point or time, get butt hurt at own risk.

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OfflineMontock
Time = $
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Posts: 1,838
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Re: Agar transfers showing super slow activity, sometimes drying out [Re: Montock]
    #17321451 - 12/02/12 03:29 PM (12 years, 1 month ago)

even rr says in the very second post everything im telling you now, your post supports what im saying, not that glad wrap works, i need to link that link to notahacker


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Looking for Wife, please pm me your picture and list of skillz.


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OfflineSma11mau5
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Registered: 08/15/12
Posts: 98
Last seen: 10 years, 7 months
Re: Agar transfers showing super slow activity, sometimes drying out [Re: Montock]
    #17324626 - 12/03/12 01:40 AM (12 years, 1 month ago)

That the glad doesn't breath perfectly shouldn't matter, as strapping bands around t leaving areas where it can breathe anyway rather than tightly wrap around the edge to create an airtight seal

Sorry just not willing to shell out so much extra for the parafilm is there no one who is successfull with glad???

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Offlineskippydude
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Re: Agar transfers showing super slow activity, sometimes drying out [Re: Sma11mau5]
    #17325403 - 12/03/12 08:11 AM (12 years, 1 month ago)

I have.

Never had drying.

Just used parafilm for the first time yesterday.

I had condensation build up on a couple dishes, never any drying out.

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OfflineSma11mau5
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Registered: 08/15/12
Posts: 98
Last seen: 10 years, 7 months
Re: Agar transfers showing super slow activity, sometimes drying out [Re: skippydude]
    #17326045 - 12/03/12 11:05 AM (12 years, 1 month ago)

Quote:

skippydude said:

Sounds like you have plain agar no nutrients.

How many grams is used per 1000ml?

If it is around 20 grams it is likely plain agar.

If it is more like 40 grams it is probably MEA.

I have been using saran wrap, the only problem I have had is with a couple dishes I refrigerated right side up, the collected a tablespoon of water in them.

http://www.shroomery.org/forums/showflat.php/Number/17200242/page/2




It was arround 56gm, it is defiantly a nutrient containing agar, if you introduce anything to the plates it will grow, just the issue I'm having trouble with is the speed of growth vs the length of time the agar can keep without drying

I guess I will go back to tightly wrapping around the edge, and do you guys prefer to store them in a fridge or at room temp when you are trying to grow out a sample? Don't think the main fridge would be a 'clean' place as we are major cheese lovers lol

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Offlineskippydude
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Re: Agar transfers showing super slow activity, sometimes drying out [Re: Sma11mau5]
    #17326990 - 12/03/12 02:28 PM (12 years, 1 month ago)

Room temp for growing and frige for storage and slower growth.

Edited by skippydude (12/03/12 02:29 PM)

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InvisibleZarotti
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Registered: 05/12/12
Posts: 314
Re: Agar transfers showing super slow activity, sometimes drying out [Re: skippydude]
    #17327367 - 12/03/12 03:32 PM (12 years, 1 month ago)

I read you can use pins for your agar work

1.Would a 6 month old air dried pin colonize on an agar plate ?

What is the maximum time until the harvested pin is
not usable for agar anymore ?

2.After how many days should you see growth after inoculating an agar dish with a spore syringe ? 68 F(syringe is the only source currently)

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OfflineMontock
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Re: Agar transfers showing super slow activity, sometimes drying out [Re: Zarotti]
    #17329289 - 12/03/12 08:51 PM (12 years, 1 month ago)

Quote:

Zarotti said:
I read you can use pins for your agar work

1.Would a 6 month old air dried pin colonize on an agar plate ?

What is the maximum time until the harvested pin is
not usable for agar anymore ?

2.After how many days should you see growth after inoculating an agar dish with a spore syringe ? 68 F(syringe is the only source currently)



something will grow

id give it 7 days or so, usually within 3-5 i can see something starting


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Looking for Wife, please pm me your picture and list of skillz.


I'm wrong all the time, my posts are subject to being wrong at any  point or time, get butt hurt at own risk.

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InvisibleSpitballJedi
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Re: Agar transfers showing super slow activity, sometimes drying out [Re: Montock]
    #17329447 - 12/03/12 09:15 PM (12 years, 1 month ago)

I would take the guessing out of it and ask the place , where you purchased the agar, to see the ingredients list.

It may not be plain agar, but it may not have the nutes you need for mycology.


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