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Offlinegrivel99
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Is this an ok agar Tek?
    #24772652 - 11/10/17 12:18 AM (3 years, 5 months ago)

Step 1 mix 12g dried malt extract(light) with 8g agar agar powder in bottle

Step 2 add 500ml water and shake

Step 3 pressure cook (20 minutes at 12psi ok)

Step 4 let cool take into sab and poor into 90mm/15mm pitri dishes (I also have 30mm/10mm pitri dishes and thought they would be ok for doing the first few jumps until I see something good in the mycelium)

Step 5 add 1/4cc of multispore syringe (b+ , there cambodian has inoculated 6 jars happily so I think they're ok) and move around with loop

Step 6 when half pitri dish is covered take a few segments move to fresh agar and Chuck leftovers in garden

Step 7 repeat until I see something good then when i have a few fully rhizomorphic? Mother plates start moving to grain (will probably be testing batches before then though


Never done this before any help appreciated thank you


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Edited by bodhisatta (11/10/17 12:23 AM)


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Re: Is this an ok agar Tek? [Re: grivel99]
    #24772666 - 11/10/17 12:27 AM (3 years, 5 months ago)

1. I would use 10g malt and 10g agar to 500mL of water

2. I would bring it to a boil. then pour into a suitable bottle to pressurecook/autoclave leaving behind any sediment that formed

3. at 12 psi I would do 30m

4. :thumbup:

5. no, add a single drop or less. use an inoculation loop, put a single drop on that, then zig zag the loop across the agar.

6. yes

7. rhizomorphic doesn't really mean shit. you just want healthy growth. anything that's clean goes to a new dish. as soon as you have clean mycelium(can be as soon as dish #2) you can move it to grain. appearance on agar has nothing to do with performance. if you lower the nutrition in agar you'll get more rhizomorphic growth.

use the search engine, don't look at old crap.

https://www.shroomery.org/forums/dosearch.php?forum%5B%5D=f2&words=agar+tek&namebox=&expert=1&replybox=&how=all&where=body&tosearch=main&newerval=3&newertype=y&olderval=&oldertype=y&minwords=&maxwords=&image=1&limit=25&sort=r&way=d&showmain=1

the first result has all the info you need.
search for agar tek, by trusted cultivators, has pictures, in main posts(because you're looking for a TEK), newer than 3-5 years

also this one too is a must read
https://www.shroomery.org/forums/showflat.php/Number/19208976


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OfflineKrombopulosM
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Re: Is this an ok agar Tek? [Re: bodhisatta]
    #24772685 - 11/10/17 12:50 AM (3 years, 5 months ago)

:whathesaid:


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Offlinegrivel99
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Re: Is this an ok agar Tek? [Re: KrombopulosM]
    #24772716 - 11/10/17 01:23 AM (3 years, 5 months ago)

Thank you..
And by 1/4cc I mean a quarter of a cc

Bring the mixture to boil in what adzactly?


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Offlinegrivel99
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Re: Is this an ok agar Tek? [Re: grivel99]
    #24772723 - 11/10/17 01:27 AM (3 years, 5 months ago)

Oh and I've read all these teks ten times over I just see all different information from different people and tried to put it together..


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OfflineKrombopulosM
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Re: Is this an ok agar Tek? [Re: grivel99]
    #24772727 - 11/10/17 01:31 AM (3 years, 5 months ago)

Quote:

grivel99 said:
Thank you..
And by 1/4cc I mean a quarter of a cc

Bring the mixture to boil in what adzactly?




I'm not a physicist but for some reason the boiling agar does not break a sealed jar I cook 1200 ml at a time half gallon jar inside my pc.
I mix it in the jar shake tf out of it and pc at 15 psi for 30 minutes.  I used to boil it in a glass bowl in the microwave and I noticed that it bubbles really bad when it boils in a non sealed container. I would cook for 5 minutes and half of it would be all over my microwave.



Edited by KrombopulosM (11/10/17 01:33 AM)


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Offlinegrivel99
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Re: Is this an ok agar Tek? [Re: KrombopulosM]
    #24772740 - 11/10/17 01:42 AM (3 years, 5 months ago)

So do I need to boil/cook my agar first or just shake it up real good an PC it for lomger? Or microwave it n shake every minute for 3 minutes? I can't figure out how to boil any other ways other than just sticking the mixture straight in a saucepan then pouring it into bottles pc i t then into dishes when it's needed


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OfflineKrombopulosM
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Re: Is this an ok agar Tek? [Re: grivel99]
    #24772767 - 11/10/17 02:17 AM (3 years, 5 months ago)

I used to boil first but I've found there is no difference between that and just putting it straight into my pc glass and shaking a little then pc. Once the pc is depressurized u will want to take the lid off and let it sit for maybe half an hour before u shake ur agar jar again. U will see a layer on the bottom. It just needs a good shake to distribute it properly. Do not shake as soon as the pc is off. The glass will break or the seal will pop. Wait 30 minutes.


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Re: Is this an ok agar Tek? [Re: KrombopulosM]
    #24772791 - 11/10/17 02:52 AM (3 years, 5 months ago)

I made some agar plates today too. Streaked with a swab.
Quote:

KrombopulosM said:
I used to boil first but I've found there is no difference between that and just putting it straight into my pc glass and shaking a little then pc. Once the pc is depressurized u will want to take the lid off and let it sit for maybe half an hour before u shake ur agar jar again. U will see a layer on the bottom. It just needs a good shake to distribute it properly. Do not shake as soon as the pc is off. The glass will break or the seal will pop. Wait 30 minutes.







After my pc comes down to atmospheric temp, I take out my flask, I don’t agitate because I want a clear plate and it’s still warm and condensation happens but it goes away

And here’s an empty plate that I streaked just now. Which is basically nothing but the clarity and colour are nice. I tasted it and very subtle. If I get some good growth I will share my recipe. It’s super easy so I hope it works


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InvisiblebodhisattaM
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Re: Is this an ok agar Tek? [Re: grivel99]
    #24773014 - 11/10/17 09:23 AM (3 years, 5 months ago)

Quote:

grivel99 said:
Thank you..
And by 1/4cc I mean a quarter of a cc

Bring the mixture to boil in what adzactly?



Yes I know what fractions are. That's way too much to put on a petri dish.

You can boil it in whatever you want.....

If you did read new teks. Hate to say it you need to re read em..


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Offlinegrivel99
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Re: Is this an ok agar Tek? [Re: bodhisatta]
    #24773060 - 11/10/17 10:00 AM (3 years, 5 months ago)

I just figured as possibly a weak spore syringe itd be best to use more (no matter how much I shake or whatever light I hold it too I can.only see a couple black specs) I also figured 1/4cc would be a miniscule amount I'm still new to this so getting used to measurements is complicated especially when there all american..

Most teks don't fully explain everything..
They explain one bit then another Tek explains another

And it seems like ive got this down so far my only problem was my measurements for agar really..

It doesn't seem necessary to boil your agar as I know ive read about plenty people not boiling it and just PC it

Quote:

KrombopulosM said:
I used to boil first but I've found there is no difference between that and just putting it straight into my pc glass and shaking a little then pc. Once the pc is depressurized u will want to take the lid off and let it sit for maybe half an hour before u shake ur agar jar again. U will see a layer on the bottom. It just needs a good shake to distribute it properly. Do not shake as soon as the pc is off. The glass will break or the seal will pop. Wait 30 minutes



So if I was to rewrite my Tek

Step 1 mix 10g dried malt extract(light) with 10g agar agar powder in bottle

Step 2 add 500ml water and shake

Step 21/2 boil like bod... Or just don't either way

Step 3 pressure cook 30 minutes at 12 psi

Step 4 let cool take into sab and poor into 90mm/15mm pitri dishes (I also have 30mm/10mm pitri dishes and thought they would be ok for doing the first few jumps until I see something good in the mycelium)

Step 5 add a miniscule drop of multispore syringe (b+ , there cambodian has inoculated 6 jars happily so I think they're ok) and move around with inoculation loop

Step 6 when half pitri dish is covered take a few segments move to fresh agar and Chuck leftovers in garden

Step 7 repeat until I see something good then when i start getting healthy mycelium move to grains

Step 8 continue with agar until I learn how to isolate certain genes



What else... What's the ambition here... I'm always seeing people happy about there plates coz they've done enough jumps and they have a fully rhizomporhic plate (mother plate) what's so good about this if all I want is healthy mycelium

I thought I was trying to take all the segments away from eachother until there was only one left.. and I'd have to test each segments growing abilities along the line aswell because the gene could.be potent but not have a good yield and visa versa.. 
Or am I just confused


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Re: Is this an ok agar Tek? [Re: grivel99]
    #24773066 - 11/10/17 10:06 AM (3 years, 5 months ago)

After pressure cooking if there's a layer you did something wrong. The agar should be completely homogeneous after sterilization.

Boiling before helps achieve this. As you can decant off any sediment that forms from heating. "Hot break protein" precipitates out and becomes insoluble.

The ambitioois a clean culture. Spores are not completely axenic. Use agar to obtain a clean culture. That you grow with.

If you want to do isolation do it in your spare time. As testing w bunch of isolates is a lot of work and not so much yield necessarily.

Take clones from your grows those clones can be saved on agar. Those clones will have good to great performance.

Most of us rock clones.

I think I need to go edit my tek a little. I was hoping those things would be clear.


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Offlinegrivel99
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Re: Is this an ok agar Tek? [Re: bodhisatta]
    #24773110 - 11/10/17 10:29 AM (3 years, 5 months ago)

Now makes sense.. thank you I have always got all ur teks up in a tab for info whenever I need it.. but this is a forum page it's for posting things and getting help...


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Offlinegrivel99
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Re: Is this an ok agar Tek? [Re: grivel99]
    #24781093 - 11/13/17 08:29 PM (3 years, 5 months ago)

Had loads of sediment in jar after boiling... And mixing.. I boiled the water and chucked the agar and lme in that mixed around until what I though was fully dissolved but when I poured into jar to go in PC there was lots of sediment in the jar is this normal or should I just re-try


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Re: Is this an ok agar Tek? [Re: grivel99]
    #24781127 - 11/13/17 08:42 PM (3 years, 5 months ago)

I would look up no pour plates. they are really good for getting your feet wet with agar


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Re: Is this an ok agar Tek? [Re: grivel99]
    #24781174 - 11/13/17 09:05 PM (3 years, 5 months ago)

Quote:

grivel99 said:
Had loads of sediment in jar after boiling... And mixing.. I boiled the water and chucked the agar and lme in that mixed around until what I though was fully dissolved but when I poured into jar to go in PC there was lots of sediment in the jar is this normal or should I just re-try




Was it powder or strips?


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Offlinegrivel99
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Re: Is this an ok agar Tek? [Re: Citizen X]
    #24781181 - 11/13/17 09:08 PM (3 years, 5 months ago)

Have already bought everything for pour Tek and don't have the money to invest in glass petri dishes have attempted once so I know I just need to do something different but I'm not sure

And it was powder


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Re: Is this an ok agar Tek? [Re: grivel99]
    #24782269 - 11/14/17 12:58 PM (3 years, 5 months ago)

Quote:

grivel99 said:
Have already bought everything for pour Tek and don't have the money to invest in glass petri dishes have attempted once so I know I just need to do something different but I'm not sure

And it was powder




There are some recipes that just don’t click. Potato flake was one that I had no luck with.

But... I have some growth on my “anything” agar. I’m so proud :smile:


Ready for my first transfer. Out of 4 plates streaked only 1 has shown growth, 2 no growth, 1 contamination


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