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LCrow
Wanderer



Registered: 12/13/20
Posts: 102
Loc: Gloucestershire
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My First Agar (MEA, Poured)
#27122047 - 01/02/21 12:52 PM (3 years, 1 month ago) |
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Hello all,
So after lurking, researching and buying some equipment in the pursuit of messing around indoors with some semilanceata I found that in general the consensus these days is agar is good stuff so I thought I'd give it a crack.
I followed bods easy af Tek for the most part; https://www.shroomery.org/forums/showflat.php/Number/21922023/fpart/1/vc/1
I just heated my flask in a pot of boiling water instead of a microwave, although in future I may go for freshly boiled water straight into the media bottle.
Anyway here's my plates, I poured a little hot so used the mug of warm water trick to sort the condensation out and will let it cool a bit more next time, but all in all I don't think they look awful for a first go


I did have an issue with some boiling over in my PC but I think that was user error in venting right after it was done (not sure what I was thinking tbh, stupid moment) so I lost some of my solution, but still had enough for 4 plates
Now to see if they contam! No mushy goodness on them yet, but if they stay contam free I'm going to try my hand at cloning to agar
To all the other noobs out there, it'd true what they say, agar isn't that scary!
EDIT: I use an electric PC that runs at 10psi, so if yours run at this or 12psi you can do agar for sure. I now run my cycle on high for 45 minutes with success.
Edited by LCrow (02/05/21 04:38 AM)
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TigerStrike
carpophore locator



Registered: 12/22/20
Posts: 315
Last seen: 1 year, 9 months
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Re: My First Agar (MEA, Poured) [Re: LCrow]
#27122061 - 01/02/21 01:01 PM (3 years, 1 month ago) |
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Agar is never scary on the first pour. It gets demotivationg if you felt like your pour was perfect and then 4 days later your plates are full of shit. Enough to make me quit years ago and only back here from over 8 year ago attempt. Don't quit unless you are a quitter then quit.
You've got a long way to go. But I see your traveling caravan and it is traveling in the correct direction!
Congrats and welcome to agar. Don't get discouraged and give up if you experience contams. About 2 weeks ago I couldn't find my jiggler and ended up hot water bathing some agar. Got a few contam plates but most were fine. Are you eyeballing a PC? What you got your eyes on?
Edited by TigerStrike (01/02/21 01:04 PM)
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LCrow
Wanderer



Registered: 12/13/20
Posts: 102
Loc: Gloucestershire
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Re: My First Agar (MEA, Poured) [Re: TigerStrike]
#27122075 - 01/02/21 01:12 PM (3 years, 1 month ago) |
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It was my first try using an SAB too, but I'm well prepared for all the disappointment I may face along the way, as you say its not the pour as such that's troubling, I just wanted anyone who was on the fence and lurking like me know that with bods easy af Tek, the technical bits at least aren't so intimidating 
I've got myself an electric one, I know there's a lot of stigma around that, but it seems to work for others, with adjusted sterilisation times so I'm hoping it'll work for me too
Thanks for the kind words, hopefully in a week or so I'll know if I have any contams or not and can go for another try if so
Edited by LCrow (01/02/21 01:31 PM)
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Frozen Jedi
nooberist


Registered: 10/17/20
Posts: 26
Last seen: 3 years, 10 days
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Re: My First Agar (MEA, Poured) [Re: LCrow]
#27122892 - 01/02/21 09:43 PM (3 years, 1 month ago) |
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congrats on your first pour, I hope it comes out well for you! I made my first pour around 2 months ago so that anxiety is still fresh. Here's the thing, if you do get contamination, you can pour again. If you noc up a plate and get contams, you can transfer away from it with equal success. the point being, any anxiety associated with pouring, transferring, or otherwise working with agar is far outweighed by the benefits it offers in the big picture, also it's a lot of fun.
good luck!
FJ
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LCrow
Wanderer



Registered: 12/13/20
Posts: 102
Loc: Gloucestershire
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Re: My First Agar (MEA, Poured) [Re: Frozen Jedi]
#27123135 - 01/03/21 02:46 AM (3 years, 1 month ago) |
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Thanks FJ, I've just checked out your threads and it looks like you've been through the mill a bit with those contams!
Still great reference material for me over the next week or two so thanks for posting those images 
It was quite nerve wracking, and not knowing how to work in an SAB I'm sure I've probably made some mistakes and may see contams because of it, but that's science for you! As you say you can always brew a fresh batch, and move healthy tissue, for now I'll let these sit for a few days to see how clean I was
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LCrow
Wanderer


Registered: 12/13/20
Posts: 102
Loc: Gloucestershire
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Re: My First Agar (MEA, Poured) [Re: LCrow]
#27125188 - 01/04/21 03:55 AM (3 years, 1 month ago) |
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Day 2 update - plates 3 and 4 are looking okay so far and are not pictured.
Pictures 3 to 5 below show plate 1 with a yellow type growth on the periphery, I believe this will be bacterial? Around 12 o'clock
Plate 2, if you can see in images 1 and 2 has something I can't quite discern, I don't know if it's water sat on the dish that will disappear in more time, trapped air or some sort of mould or something? Its hard to even tell if it's on or under the agar! Again it is around the periphery and at around 11 o'clock in picture 2 (between and left of the reflections of my light bulbs) and 12 o'clock on picture 1
Anyhow this is my progress, I'll probably post another update in a couple of days to see if there's anything more obvious on plate 2 and whether plates 3 and 4 still look clean
Edited by LCrow (01/04/21 04:02 AM)
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LCrow
Wanderer


Registered: 12/13/20
Posts: 102
Loc: Gloucestershire
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Re: My First Agar (MEA, Poured) [Re: LCrow]
#27137815 - 01/09/21 02:51 PM (3 years, 1 month ago) |
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Pictures as at day 7
Dish 2 showing contamination, some of the yellow bacteria seen on Dish 1 as well as some slimey, clear sort of contam too
 Dish 3, with the mould growing further

Dish 2 from the front to better show the clear looking contam
Dish 4, still looking clean

Dish 1 showing the growth progress of the contam on this one

Another shot of dish 4, I assume this dish is clean and I'm pleased with one out of the four being clean. I did pour this one last and on this first pour I did have the dishes open too long in the SAB. I Poured 6 more today, with much less faffing and better sterile process so I'm hoping I'll see a better success rate there.
In the meantime this dish may well become the recipient of my first piece of tissue to attempt to clone
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LCrow
Wanderer


Registered: 12/13/20
Posts: 102
Loc: Gloucestershire
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Re: My First Agar (MEA, Poured) [Re: LCrow]
#27158716 - 01/20/21 03:39 AM (3 years, 27 days ago) |
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In the interest of completeness I'm going to post the final picture of these dishes before I toss them.
Plates 1 through 3 were contam from the start, plate 4 I did a real messy job of putting the clone tissue to the plate as per my journal. It was suggested I try hot pouring, but the plate is so nasty I don't want to be opening it in my SAB if I don't have to.
These 4 plates will all be tossed, but it was a great experience.
Plates 5 to 10 are all looking clean, 2 have tissue transfers on, which I am tracking in my journal and also have a question posted about in the getting started forum as there is fine growth but I am unsure if it is myc as I've not seen anything like it in pictures here so far
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