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Drshroomy88
The doctor
Registered: 11/17/12
Posts: 318
Loc: The WWW
Last seen: 11 months, 19 hours
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Agar/light malt extract petri dish prep?
#17512145 - 01/07/13 01:39 PM (11 years, 2 months ago) |
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So my little ones are getting close to the point where I need to prepare my agar agar and light malt extract for a petri dish culture. I know that I need to cut a sliver from the inside of an ideal specimen and drop it on the petri dish with the agar/light malt extract culture.
My question is, how do I prepare this culture, and what amounts of agar and light malt extract do I use?
I've watched this video, but it's not entirely helpful
http://www.mushroomvideos.com/Agar-Petri-Dishes
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toxetel
׀ǝʇǝxoʇ ●
Registered: 08/13/12
Posts: 708
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Re: Agar/light malt extract petri dish prep? [Re: Drshroomy88]
#17512228 - 01/07/13 01:57 PM (11 years, 2 months ago) |
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Sillyputty67
Registered: 10/06/12
Posts: 2,239
Loc: Netherlands
Last seen: 9 years, 6 months
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Re: Agar/light malt extract petri dish prep? [Re: toxetel]
#17512242 - 01/07/13 02:02 PM (11 years, 2 months ago) |
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I like to use a mason jar with a sfd. The mason jar has graduated marks on the side.
I only like to do about 7 or 8 petri dishes at a time.
So i use: 250 ML hot tap water 5 grams light dry malt extract 5 grams agar pressure cook for 30 min.
Works every time.
-------------------- 1) Everything I ever posted or say is a lie.
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MoGrow
Lysergic Funeral Procession
Registered: 11/16/12
Posts: 1,356
Loc: Da Sunshine State
Last seen: 1 year, 4 months
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Re: Agar/light malt extract petri dish prep? [Re: Drshroomy88]
#17512323 - 01/07/13 02:20 PM (11 years, 2 months ago) |
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Quote:
Drshroomy88 said: So my little ones are getting close to the point where I need to prepare my agar agar and light malt extract for a petri dish culture. I know that I need to cut a sliver from the inside of an ideal specimen and drop it on the petri dish with the agar/light malt extract culture.
My question is, how do I prepare this culture, and what amounts of agar and light malt extract do I use?
I've watched this video, but it's not entirely helpful
http://www.mushroomvideos.com/Agar-Petri-Dishes
I use 1:1 5gms Agar / 5gms DLME > 340ml of water. RR said somewhere that less neuts = more rhizo so my last batch for transfers was 5gms Agar / 4gms DLME and it looks good.
-------------------- Trades Current Contest! My current mono (pics) Ghetto Bulk Dehydrator Tek How I Made My Alcohol Lamp Oeric McKenna said: "My innoculant is heavily contaminated with mushroom mycelium"
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Drshroomy88
The doctor
Registered: 11/17/12
Posts: 318
Loc: The WWW
Last seen: 11 months, 19 hours
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Re: Agar/light <nobr><a id="FALINK_1_0_0" class="FAtxtL" href="#">malt extract</a></nobr> petri dish prep? [Re: Sillyputty67]
#17512328 - 01/07/13 02:21 PM (11 years, 2 months ago) |
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Quote:
malicom said: I like to use a mason jar with a sfd. The mason jar has graduated marks on the side.
I only like to do about 7 or 8 petri dishes at a time.
So i use: 250 ML hot tap water 5 grams light dry malt extract 5 grams agar pressure cook for 30 min.
Works every time.
Ok, at 15 or 30psi? Then you let it cool for a bit? How long? Or do you pour it straight into the petris? And do your petris need to be refrigerated after you add the piece of shroom? Or do they just stand in room temp? Light or dark room?
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Drshroomy88
The doctor
Registered: 11/17/12
Posts: 318
Loc: The WWW
Last seen: 11 months, 19 hours
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Re: Agar/light <nobr><a id="FALINK_1_0_0" class="FAtxtL" href="#">malt extract</a></nobr> petri dish prep? [Re: MoGrow]
#17512338 - 01/07/13 02:24 PM (11 years, 2 months ago) |
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Quote:
MoGrow said:
Quote:
Drshroomy88 said: So my little ones are getting close to the point where I need to prepare my agar agar and light malt extract for a petri dish culture. I know that I need to cut a sliver from the inside of an ideal specimen and drop it on the petri dish with the agar/light malt extract culture.
My question is, how do I prepare this culture, and what amounts of agar and light malt extract do I use?
I've watched this video, but it's not entirely helpful
http://www.mushroomvideos.com/Agar-Petri-Dishes
I use 1:1 5gms Agar / 5gms DLME > 340ml of water. RR said somewhere that less neuts = more rhizo so my last batch for transfers was 5gms Agar / 4gms DLME and it looks good.
Ok, sounds legit, and what do you pour it into when you do the PC? And at what PSI for how long?
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PussyFart
Retired Cultivation Extrodinaire
Registered: 04/08/12
Posts: 22,502
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Re: Agar/light <nobr><a id="FALINK_1_0_0" class="FAtxtL" href="#">malt extract</a></nobr> petri dish prep? [Re: Drshroomy88]
#17512342 - 01/07/13 02:24 PM (11 years, 2 months ago) |
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I suggest you fill the PC with water until half of the agar jar is submerged, this will help prevent boil over.
15psi for 30-40 minutes and then let it sit in the PC for 2 hours...
Take the agar out and let it cool to the point where you need no heat protection to pour it, then pour the dishes.
Waiting for it to cool like this will almost completely eliminate condensation and excess moisture in the dishes.
-------------------- THIS HOBBY IS NOT FOR THE IMPATIENT! PLEASE BE PATIENT, DON'T BE A PATIENT! A Tale of 10 Isolates, GT Cluster Clone Monotubs, RR's Let's Grow Mushrooms DVD, SGFC(Shotgun Fruiting Chamber), Monotub Tek, Damion5050's Coir Tek, TL's Tek List, Frank's Tek List, EvilMushroom666's Pasteurization Tek, How It Should & Shouldn't Look - NEW CULTIVATORS GUIDE *** *** AFGHAN KUSH GROW LOG *** ***
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Sillyputty67
Registered: 10/06/12
Posts: 2,239
Loc: Netherlands
Last seen: 9 years, 6 months
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Re: Agar/light <nobr><a id="FALINK_1_0_0" class="FAtxtL" href="#">malt extract</a></nobr> petri dish prep? [Re: Drshroomy88]
#17512368 - 01/07/13 02:29 PM (11 years, 2 months ago) |
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Generally everything referring to a PC cycle, we are all referring to 15 psi as the standard. I also read about less nutrient/more rhizo. My thinking is you want the standard 3% MEA for germinating spores, or like Mogrow said, which is probably closer to 2% MEA for transfers.
-------------------- 1) Everything I ever posted or say is a lie.
Edited by Sillyputty67 (01/07/13 02:40 PM)
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Drshroomy88
The doctor
Registered: 11/17/12
Posts: 318
Loc: The WWW
Last seen: 11 months, 19 hours
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Re: Agar/light <nobr><a id="FALINK_1_0_0" class="FAtxtL" href="#">malt extract</a></nobr> petri dish prep? [Re: Sillyputty67]
#17512414 - 01/07/13 02:39 PM (11 years, 2 months ago) |
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Quote:
Notahacker420 said: I suggest you fill the PC with water until half of the agar jar is submerged, this will help prevent boil over.
15psi for 30-40 minutes and then let it sit in the PC for 2 hours...
Take the agar out and let it cool to the point where you need no heat protection to pour it, then pour the dishes.
Waiting for it to cool like this will almost completely eliminate condensation and excess moisture in the dishes.
Quote:
malicom said: Generally everything referring to a PC cycle, we are all referring to 15 psi as the standard. I also read about less nutrient/more rhizo. My thinking is you want the standard 3% MEA for germinating spores, or like Mogrow said, which is probably closer to 2% MEA for transfers.
Ok thanks guys, I think I got it!
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MoGrow
Lysergic Funeral Procession
Registered: 11/16/12
Posts: 1,356
Loc: Da Sunshine State
Last seen: 1 year, 4 months
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Re: Agar/light <nobr><a id="FALINK_1_0_0" class="FAtxtL" href="#">malt extract</a></nobr> petri dish prep? [Re: Drshroomy88]
#17512502 - 01/07/13 02:54 PM (11 years, 2 months ago) |
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Quote:
Drshroomy88 said:
Quote:
MoGrow said:
Quote:
Drshroomy88 said: So my little ones are getting close to the point where I need to prepare my agar agar and light malt extract for a petri dish culture. I know that I need to cut a sliver from the inside of an ideal specimen and drop it on the petri dish with the agar/light malt extract culture.
My question is, how do I prepare this culture, and what amounts of agar and light malt extract do I use?
I've watched this video, but it's not entirely helpful
http://www.mushroomvideos.com/Agar-Petri-Dishes
I use 1:1 5gms Agar / 5gms DLME > 340ml of water. RR said somewhere that less neuts = more rhizo so my last batch for transfers was 5gms Agar / 4gms DLME and it looks good.
Ok, sounds legit, and what do you pour it into when you do the PC? And at what PSI for how long?
i use no pour agar tek.
Transfered 3 days ago using 4/5 receipe.
yes everything here is 15Psi and same as NAH420 stated 30 min. with no pour boil over isnt an issue.
That being said , once i get a strong understanding of all things agar and isolation, ill be moving over to dishes, but being single use, i started with jars.
-------------------- Trades Current Contest! My current mono (pics) Ghetto Bulk Dehydrator Tek How I Made My Alcohol Lamp Oeric McKenna said: "My innoculant is heavily contaminated with mushroom mycelium"
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PussyFart
Retired Cultivation Extrodinaire
Registered: 04/08/12
Posts: 22,502
Loc: Orbiting Earth
Last seen: 2 months, 14 days
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Re: Agar/light <nobr><a id="FALINK_1_0_0" class="FAtxtL" href="#">malt extract</a></nobr> petri dish prep? [Re: MoGrow]
#17512514 - 01/07/13 02:56 PM (11 years, 2 months ago) |
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Quote:
MoGrow said: i use no pour agar tek.
Transfered 3 days ago using 4/5 receipe.
yes everything here is 15Psi and same as NAH420 stated 30 min. with no pour boil over isnt an issue.
That being said , once i get a strong understanding of all things agar and isolation, ill be moving over to dishes, but being single use, i started with jars.
Are you storing your dishes upside down?
Or did you just do that to take a pic?
-------------------- THIS HOBBY IS NOT FOR THE IMPATIENT! PLEASE BE PATIENT, DON'T BE A PATIENT! A Tale of 10 Isolates, GT Cluster Clone Monotubs, RR's Let's Grow Mushrooms DVD, SGFC(Shotgun Fruiting Chamber), Monotub Tek, Damion5050's Coir Tek, TL's Tek List, Frank's Tek List, EvilMushroom666's Pasteurization Tek, How It Should & Shouldn't Look - NEW CULTIVATORS GUIDE *** *** AFGHAN KUSH GROW LOG *** ***
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MoGrow
Lysergic Funeral Procession
Registered: 11/16/12
Posts: 1,356
Loc: Da Sunshine State
Last seen: 1 year, 4 months
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Re: Agar/light <nobr><a id="FALINK_1_0_0" class="FAtxtL" href="#">malt extract</a></nobr> petri dish prep? [Re: PussyFart]
#17512536 - 01/07/13 03:00 PM (11 years, 2 months ago) |
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Just to take a pic. The bad thing about jars is you can only pic one side and the camera wants to focus on the lid not the culture, dishes are going to be nice all around just waiting to get agar down. first couple runs i was fighting mold contams, my sterile and SAB tek is good now i never see mold but im fighting a bacterial transfer issue. i got 4:5 clean on the last transfer, so i think ive got that taken care of too. cant wait to start using petris. these 4 stay clean and ill be using them next transfer. Then i get to deal with prepping agar the proper way.
-------------------- Trades Current Contest! My current mono (pics) Ghetto Bulk Dehydrator Tek How I Made My Alcohol Lamp Oeric McKenna said: "My innoculant is heavily contaminated with mushroom mycelium"
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Sillyputty67
Registered: 10/06/12
Posts: 2,239
Loc: Netherlands
Last seen: 9 years, 6 months
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Re: Agar/light <nobr><a id="FALINK_1_0_0" class="FAtxtL" href="#">malt extract</a></nobr> petri dish prep? [Re: MoGrow]
#17512597 - 01/07/13 03:12 PM (11 years, 2 months ago) |
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Nice rhizo's Mo.
how hard is it to cut a transfer from the bottom of the jar?
-------------------- 1) Everything I ever posted or say is a lie.
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MoGrow
Lysergic Funeral Procession
Registered: 11/16/12
Posts: 1,356
Loc: Da Sunshine State
Last seen: 1 year, 4 months
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Re: Agar/light <nobr><a id="FALINK_1_0_0" class="FAtxtL" href="#">malt extract</a></nobr> petri dish prep? [Re: Sillyputty67]
#17512630 - 01/07/13 03:18 PM (11 years, 2 months ago) |
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Thx.
Its easy i use an Xacto knife its 5.5" from tip to tail and the jar with lid is 2.5" i havent had any issues only issue ive run into is when i flame sterlize my blade the tip is so thin it curls but i got the knife that seats the fine blade cant remember the number but i need to get a medium blade knife, in the meantime i snipped the tip of the blades with cutters, anyways its an issue because its hard to get the transfer off the blade even swiping down into the fresh agar. I dont transfer from the bottom of the jar tho i take my agar about 1/2 way through and "pop" it out for lack of a better description, ill post a pic next time i transfer.
-------------------- Trades Current Contest! My current mono (pics) Ghetto Bulk Dehydrator Tek How I Made My Alcohol Lamp Oeric McKenna said: "My innoculant is heavily contaminated with mushroom mycelium"
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Drshroomy88
The doctor
Registered: 11/17/12
Posts: 318
Loc: The WWW
Last seen: 11 months, 19 hours
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Re: Agar/light <nobr><a id="FALINK_1_0_0" class="FAtxtL" href="#">malt extract</a></nobr> petri dish prep? [Re: Sillyputty67]
#17512749 - 01/07/13 03:40 PM (11 years, 2 months ago) |
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Quote:
malicom said: Nice rhizo's Mo.
how hard is it to cut a transfer from the bottom of the jar?
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