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Offlinemycopiss
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First Time doing AGAR
    #26178350 - 09/09/19 08:44 PM (1 year, 5 months ago)

This will be my first attempt at doing AGAR work. Would it be better to use spore prints to agar or small pieces of mushroom, cut from inside the mushroom?

What is the difference? Pros and Cons of each?

I purchased some pre made agar plates. Next time I will attempt to make the agar plates myself.

Any other tips feel free to let me know! Im sure It probably wont go too great being my first attempt, but I will give it my best shot.

I keep seeing photos of these huge flushes with full coverage.. I want to make it happen =)


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Offlinejcm4620
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Re: First Time doing AGAR [Re: mycopiss]
    #26178500 - 09/09/19 09:56 PM (1 year, 5 months ago)

read and read then read sum more b4 u try anything. u will have a better chance if you find a tek and follow it. start by looking up the terms used so you understand what it is that is being described in the teks and then when you have a question you will b able to ask it in a way that will help us help u better. there is alot of good info on here put together by ppl who have much knowledge so take advantage of the search engine here it is the best thing on the site. i promise you that it contains everything ul ever need happy growing


--------------------
PANAEOLUS FRUITING MADE SIMPLE



Edited by jcm4620 (09/09/19 09:57 PM)


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Offlinemycopiss
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Re: First Time doing AGAR [Re: jcm4620]
    #26178532 - 09/09/19 10:10 PM (1 year, 5 months ago)

Are there any particular Teks you recommend for somebody just getting started?

I definitely plan on staring at the computer for hours reading as much as possible!


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OfflineCthulhu23
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Re: First Time doing AGAR [Re: jcm4620] * 1
    #26178535 - 09/09/19 10:11 PM (1 year, 5 months ago)

Why not do both?  Take a clone, AND start from spores.  They're both great learning experiences.

The difference between doing a swipe from a spore print and taking a clone is that with the spore print you'll get a significant variation in genetics and therefore the characteristics of the organism will be varied.  With a clone (taking a piece of the fruit to agar), you'll get less variability.  In essence, if you take a clone, you're automatically selecting for certain traits (given that your environmental conditions - such as sub moisture, surface conditions, FAE, etc are all consistent).  From spore print, you'll also learn what new mycelial growth looks like.  In both instances, you'll learn how to clean your sample by transferring a clean area of the original plate to a new plate ("cleaning").

I'd suggest a few things.  First, read through the known/trusted teks you can find.  Bod has a great page with a ton of links listed for teks and other useful info; and especially for you, an agar tek.  Become very friendly with the search engine (Bod has a tek for that!) and start using it to look at threads about cloning vs MS, etc.

The flushes with full coverage have a few things going for them (mind you, I'm aiming for that myself - I've gotten close!).  They're from people who usually have MANY grows under their belts.  These guys have had tons of crap tubs, I'm sure of it.  They also have their conditions (FAE, temp, moisture) dialed in for the tubs and strains they are using.  And - they have likely taken clones of strains that have produced said canopies under the dialed in conditions!

Good luck!


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OfflineCthulhu23
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Re: First Time doing AGAR [Re: Cthulhu23]
    #26178537 - 09/09/19 10:11 PM (1 year, 5 months ago)



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Offlinejcm4620
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Re: First Time doing AGAR [Re: Cthulhu23]
    #26178605 - 09/09/19 10:41 PM (1 year, 5 months ago)

lol u beat me to it that tek is great to start with for sure


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PANAEOLUS FRUITING MADE SIMPLE



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OfflineNesmaster01
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Re: First Time doing AGAR [Re: jcm4620]
    #26178676 - 09/09/19 11:25 PM (1 year, 5 months ago)

Interactive Agar

This is the thread that literally got me hooked on agar. We should do this sort of thing on here again. It was super helpful for me. Would be a good read if you want.


--------------------
"Life is really simple, but we insist on making it complicated."
“I hear and I forget. I see and I remember. I do and I understand.”
“When it is obvious that the goals cannot be reached, don’t adjust the goals, adjust the action steps.”
– Confucius


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Offlinemycopiss
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Re: First Time doing AGAR [Re: Nesmaster01]
    #26179182 - 09/10/19 09:26 AM (1 year, 5 months ago)

so to get the full coverage and bigger yield, are most using the clone method vs spores to agar?
Or are they about equal as far as yield goes?

I have 4 strains I want to clone. Should I start with 2 plates of each, just incase some of them get contaminated?


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OfflineNesmaster01
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Re: First Time doing AGAR [Re: mycopiss]
    #26179310 - 09/10/19 11:07 AM (1 year, 5 months ago)

So this is a little bit of a multi faced question but I will attempt to answer.

The purpose of spores 2 Agar is to ensure a clean sample is going to the grain. Spores are added to the agar and I usually take about 5-6 plates from each MS agar. I then take as many transfers as needed to get completely clean agar samples to put to grain.

The purpose of cloning 2 agar is to help isolate the traits from the fruits for the next fruits. You take a clone from a large fruit or cluster from the center of the body between the stipe and cap. This will still contain multiple genetics in a fruiting body.

I take multiple plates of everything that I do, my purpose is that so I can cherry pick the best of the best. I also label my plates with something that I will understand in the future and put plates I may want to use again in my cold storage in case I need to take a step back.

Hope this helps to answer your questions.


--------------------
"Life is really simple, but we insist on making it complicated."
“I hear and I forget. I see and I remember. I do and I understand.”
“When it is obvious that the goals cannot be reached, don’t adjust the goals, adjust the action steps.”
– Confucius


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Offlinemycopiss
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Re: First Time doing AGAR [Re: Nesmaster01]
    #26186425 - 09/13/19 07:17 PM (1 year, 5 months ago)

So once I use a mushroom and split it in half, can I re-use that same mushroom again for more cloning and save it in the fridge? or do I have to toss it?

Also how long does the petri dish take to fully colonize?
How many transfers on average do you guys do to get a clean sample?

And is there any benefit to running a hepa filter in the room while doing agar? or


Edited by mycopiss (09/13/19 07:45 PM)


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OfflineCthulhu23
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Re: First Time doing AGAR [Re: mycopiss] * 1
    #26186773 - 09/13/19 10:11 PM (1 year, 5 months ago)

Quote:

mycopiss said:
So once I use a mushroom and split it in half, can I re-use that same mushroom again for more cloning and save it in the fridge? or do I have to toss it?

Also how long does the petri dish take to fully colonize?
How many transfers on average do you guys do to get a clean sample?

And is there any benefit to running a hepa filter in the room while doing agar? or




The reason you split the shroom open to take a sample for clones is because the exterior is contaminated while the interior is not (ostensibly).  So by storing the shroom you're just asking for more contamination.  Take as many samples as you need, then eat/dry for later.

A petri dish takes as long as it takes to colonize.  It just depends.  Some colonize quickly, some more slowly.  I have t1 PESA that’s been on Agar for 2-3 weeks and is about 3/4 colonized (dirty/bad agar I’m thinking cuz to me that’s slow).

I can get good looking plates at t3 and t4, but I’m not as anal as others.

There is no advantage to running a heap filter unless it’s part of a flow hood.  I’d actually be more concerned with the filter kicking up air currents while doing agar work than cleaning the air.  Air currents = bad especially if near your SAB.


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OfflineNesmaster01
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Re: First Time doing AGAR [Re: Cthulhu23]
    #26186929 - 09/13/19 11:34 PM (1 year, 5 months ago)

Solid and sound advice IMHO.


--------------------
"Life is really simple, but we insist on making it complicated."
“I hear and I forget. I see and I remember. I do and I understand.”
“When it is obvious that the goals cannot be reached, don’t adjust the goals, adjust the action steps.”
– Confucius


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Offlinemycopiss
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Re: First Time doing AGAR [Re: Nesmaster01]
    #26187646 - 09/14/19 10:51 AM (1 year, 5 months ago)

It seems the first attempt is never 100% clean right?

I've had these mushrooms I wanted to clone stored in the fridge for over a week now, are they still safe to use for agar plates?

Does it make a difference if I use a butane torch to sterilize the knife or do I absolutely have to get an alcohol lamp?
Would I have to leave the torch running the whole time while doing agar work?  Will turning it off and lighting it stir up too much air?

I'm nervous about screwing this up, but I know it will pay off in the end. Trial and error!


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OfflineReliableSporesV
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Re: First Time doing AGAR [Re: Cthulhu23]
    #26187666 - 09/14/19 11:03 AM (1 year, 5 months ago)

Great post.


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Re: First Time doing AGAR [Re: Nesmaster01]
    #26187669 - 09/14/19 11:05 AM (1 year, 5 months ago)

Agar really is the best.  Especially for contamination control.


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Re: First Time doing AGAR [Re: Cthulhu23]
    #26187672 - 09/14/19 11:06 AM (1 year, 5 months ago)

Excellent post.


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Offlinefootpath
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Re: First Time doing AGAR [Re: ReliableSpores]
    #26187740 - 09/14/19 11:47 AM (1 year, 5 months ago)

You can use any flame whatsoever, so long as it can heat your instrument to the point of incandescence. Natural gas and propane torches are preferred.

You don't have to leave it running the whole time, but it takes steps out of the process.

Since your torch is outside of your still air box, you wont be kicking up the still air that matters, unless you were to point your torch at the arm holes.


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OfflineNesmaster01
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Re: First Time doing AGAR [Re: mycopiss]
    #26187758 - 09/14/19 11:58 AM (1 year, 5 months ago)

The fresher they are the higher the chance of success, that being said store bought mushrooms are cloned all of the time and a week isn’t an unheard of occurrence.


--------------------
"Life is really simple, but we insist on making it complicated."
“I hear and I forget. I see and I remember. I do and I understand.”
“When it is obvious that the goals cannot be reached, don’t adjust the goals, adjust the action steps.”
– Confucius


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OfflineCthulhu23
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Re: First Time doing AGAR [Re: Nesmaster01]
    #26188335 - 09/14/19 05:34 PM (1 year, 5 months ago)

What the last two posts said is spot on.

And yeah, first transfer is usually not 100% clean but...it’s a numbers game.  You CAN spawn from a t1, but it’s risky.  I’ve done it (g2a) and I regret it - multiple batches of grain, into the compost pile.


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Offlinemycopiss
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Re: First Time doing AGAR [Re: Cthulhu23]
    #26189491 - 09/15/19 12:30 PM (1 year, 5 months ago)

If I use a cookie tray in the bottom of the SAB do I simply wipe with alcohol to clean it?

Also, how much soap do I put in my water spray bottle?

Is the SAB technique the same used with AGAR, when using the SAB for G2G or Innoculating Grains? Wipe all supplies with alcohol, spray inside of box with soapy water, damp towel on bottom etc.?

With grain2grain or innoculating in SAB,  do I set them on the damp towel or should I use a cookie tray to place them on well? Or is a clean plastic table surface ok

I am also assuming running a hepa air purifier during G2G or inoculation in SAB is not good? What about running it in the room for 1 hr before doing SAB work , then shutting it off?


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Mushrooms, Mycology and Psychedelics >> Mushroom Cultivation

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