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OfflineBandboy
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Re: BOD's Comprehensive Agar TEK [Re: Kimble]
    #25178785 - 05/01/18 07:53 PM (5 years, 8 months ago)

Any particular agar formula best suited for Atl#7?


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InvisibleNobler Hino
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Re: BOD's Comprehensive Agar TEK [Re: Bandboy]
    #25178830 - 05/01/18 08:16 PM (5 years, 8 months ago)

lmea works great (light malt extract agar)


--------------------

"The sacred mushroom takes me by the hand and brings me to the world where everything is known. It is they, the sacred mushrooms, that speak in a way I can understand. I ask them and they answer me.”
– Maria Sabina


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OfflineBandboy
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Re: BOD's Comprehensive Agar TEK [Re: Nobler Hino]
    #25178896 - 05/01/18 09:00 PM (5 years, 8 months ago)

Thx.


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OfflineLegend21
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Re: BOD's Comprehensive Agar TEK [Re: Nobler Hino] * 1
    #25257453 - 06/09/18 06:59 AM (5 years, 7 months ago)

I’m a little confused with the water level after researching a bit. Bod you say you like to have the water level up to the level of agar when in the PC to prevent boiling over but I’ve seen other posts where it says not to let the water touch the media bottles or ones just recommending the normal water level for your pc and raising the trivet. I’m guessing all of these methods work but if you put the water up to the level of agar wouldn’t it just be boiling and not steam sterilizing?

I just got my 500ml media bottle and am about to make some agar so I didn’t want to screw it up.


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Offlineverum subsequentis
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Re: BOD's Comprehensive Agar TEK [Re: Legend21]
    #25257764 - 06/09/18 10:50 AM (5 years, 7 months ago)

i don't know about not filling it up to the level of the agar because i always do. but i can tell you that it will still sterilize it sufficiently. i've poured thousands of plates after having sterilized with the water up to the agar level and it's always fine.


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InvisiblebodhisattaMDiscordReddit
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Re: BOD's Comprehensive Agar TEK [Re: verum subsequentis]
    #25257775 - 06/09/18 11:01 AM (5 years, 7 months ago)

Steam transfers heat in the sterilizer but liquids are very easy to sterilize and the cap is loose anyway.

The reason for the added water is thermal mass. So it cools down slower so your agar doesn't boil over.

If your agar is 245F but the water around it has cooled to 240F it will start boiling. The less water in your cooker the faster it cools and may cool faster than the agar.

Follow the directions for agar here for agar. And follow the directions in my PC tek for jars of grain and etc..


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OfflineLegend21
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Re: BOD's Comprehensive Agar TEK [Re: bodhisatta]
    #25257838 - 06/09/18 11:48 AM (5 years, 7 months ago)

Awesome thanks guys. What I did this time is just put it half way up where the agar level was. Then followed the rest of the Tek to a T and have some crystal clear Petris!

Thanks again!


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OfflineWolfenstein23
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Re: BOD's Comprehensive Agar TEK [Re: Legend21]
    #25261402 - 06/11/18 11:56 AM (5 years, 7 months ago)

That's a pretty plate you got there, friend :smile: Hope mine come out half as good.

I'm currently trying to nail down what recipe I want to start with. Grabbing some of my "lab" equip. outta storage today and hopefully getting this show on the road shortly.

To BOD! Thank you so much for the plethora of information. I greatly enjoy pouring over your work, as always.


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OfflineNunyabiz
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Re: BOD's Comprehensive Agar TEK [Re: bodhisatta]
    #25281612 - 06/20/18 06:10 PM (5 years, 7 months ago)

When inoculating petris, is there an ideal? Is it better to do a drop in the middle in hope of a getting growth outwards that you can put into spawn jars immediately after maybe taking some transfers from the outer edges? Or is it better/ faster to streak the plates and isolate a potent growth then transferring? Assuming both plates are clear, isn't it faster to do the drop in the center from a syringe or spinning a sterile swab in the center from a spore print? Thanks in advance.

Thanks for all the great teks!


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Invisiblehermanium

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Re: BOD's Comprehensive Agar TEK [Re: bodhisatta]
    #25284949 - 06/22/18 10:31 AM (5 years, 7 months ago)

If doing MS to agar and the culture  appears free from contaminants the first round, is it still recommended to do a transfer? I can’t tell if your suggestion to do a transfer is to ensure a clean culture or select for healthy genetics, or both.


Edited by hermanium (06/22/18 10:33 AM)


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Offlineverum subsequentis
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Re: BOD's Comprehensive Agar TEK [Re: hermanium]
    #25284982 - 06/22/18 10:51 AM (5 years, 7 months ago)

i'd advise another transfer or two if you are a noob and can't absolutely know what you are looking at. it's just risky to put your germ plate to grain.


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Offlinekerberus
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Re: BOD's Comprehensive Agar TEK [Re: bodhisatta]
    #25303849 - 07/02/18 05:30 AM (5 years, 6 months ago)

Hi Bod/guys, i have followed this tek and poured 20 plates and then bagged them up in individual food bags. due to various reasons i didnt get to put some spores on till a couple of months later.
When i got them out 2 of the plates had contaminated but the rest looked fine so i went ahead and put a drop of B+ from a syringe off one of our sponsors (2 drops on one plate, I'm still learning!) on to 6 plates.
5 days later 4 of the plates show no sign of anything and 2 of the plates including the one with 2 drops on have the growth shown in the pictures.

This is the 2 drop plate, the other plate has the same sort of growth but more uniform.


my questions are;
The ones with no growth, could this be because the agar is too dry?
Do the 2 plates look like healthy growth?
If healthy should i transfer now or wait a few more days?
Can i use the petri dishes with no growth if the agar is fine to transfer to or would you recommend i pour new dishes for subsequent transfers?
Thanks for all the info Bod, i am happy that i took your advice and went straight to agar!

Kerberus
PS. is there a tek on how to take good petri dish photos!


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InvisibleMadHatter333
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Re: BOD's Comprehensive Agar TEK [Re: kerberus]
    #25303982 - 07/02/18 07:38 AM (5 years, 6 months ago)

So I don’t own a microwave (I know.. big shock, lol) is there anyway I can go a different route in the microwave parts? I want to work with dishes soon instead of plates for clarity. This is all great info Bod!


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InvisiblebodhisattaMDiscordReddit
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Re: BOD's Comprehensive Agar TEK [Re: MadHatter333]
    #25304134 - 07/02/18 09:13 AM (5 years, 6 months ago)

You can put it in boiling water just takes forever.


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InvisibleMadHatter333
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Re: BOD's Comprehensive Agar TEK [Re: bodhisatta]
    #25304170 - 07/02/18 09:29 AM (5 years, 6 months ago)

How long would you say? Do I need the agar to start boiling inside the bottle?


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Offlineverum subsequentis
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Re: BOD's Comprehensive Agar TEK [Re: MadHatter333]
    #25304641 - 07/02/18 02:41 PM (5 years, 6 months ago)

kerberus,

your first photos look a little concerning but honestly the picture is too shitty for me to be sure. your second photo looks pretty freaking good for ms dropped to agar. i'd take a transfer or two now. there is no reason to wait longer.

you could use the plates that didn't grow anything but it's not a good idea for two reasons.

1: you aren't being patient enough. spores don't always take off quickly. it could pop to life in a month or it could take a few days.

2: just because you don't see something growing (yet) doesn't mean it isn't. some contams (specially bacteria) can be quite sneaky.

you should probably just pour new plates.

as far as photos go, i don't know of a tek but i like front lit with a black background when i want to see morphological detail and backlit photos when trying to identify sectors. it helps to take pictures as close up as possible, straight on (not from an angle) and be sure that you are focused on the culture as opposed to the plastic or condensation.


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Offlinekerberus
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Re: BOD's Comprehensive Agar TEK [Re: verum subsequentis]
    #25305688 - 07/03/18 02:27 AM (5 years, 6 months ago)

Cheers verum,
just about to transfer now and will not re-use plates!

I will do this plate first and see if i can get a better shot of the other one.

Thank you for the reply.

Edit. 3 hours later and transferring to new plates is not as easy as i was expecting! pretty certain 2 of the 6 plates have no transferred material and look like Freddy Kruger has had his hand in it.
On a positive note poured these in the hope that i haven't totally f-up the original plate and the 6 new ones.



Edited by kerberus (07/03/18 06:18 AM)


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OfflineCultivator3000
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Re: pour agar TEK [Re: kerberus]
    #25307880 - 07/04/18 09:11 AM (5 years, 6 months ago)

Thanks for writing this. I enjoyed the read and thank you for the knowledge. :tongue2:


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OfflineSoulsearchin2018
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Re: pour agar TEK [Re: Cultivator3000]
    #25307920 - 07/04/18 09:41 AM (5 years, 6 months ago)

Hey guys, thanks for all the knowledge shared for everyone. I've jumped in head first and have so many different projects going on right now.

In regards to this thread though I do have my first 2 sets of plates of Agar and I would appreciate some advise on what Im looking at here and next steps.

Here are some plates I put together, please excuse the very rookie parafilm application:

First set of plates,



Current status:



Those are one strain/variety and here are a second set I put together:



And the current status:




My question is, what should I be looking for to cut out and transfer to another plate? I made pasty plates with the little tupper ware totes and I didn't really like how they turned out (I didn't wrap in foil) to messy and watery inside. I will probably try BRF agar in half pint jars next.

But again in regards to these plates what do you guys suggest I do from here?


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OfflineLegend21
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Re: pour agar TEK [Re: Soulsearchin2018]
    #25307940 - 07/04/18 10:02 AM (5 years, 6 months ago)

I would transfer all of those as soon as possible. Are they from a spore syringe? Usually when germinating spores you want to do the first transfer before it grows out too much and take a small piece from the leading edge.


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