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Home Grower
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How many agar dishes?
#28184798 - 02/13/23 12:51 AM (1 year, 10 months ago) |
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Hi, I intend to sprinkle some spores over agar and then, after the agar gets colonized, I am going to do a clean up by transferring the healthy pieces of mycelium to other agar dishes.
My question is: how many dishes should I initially inoculate with spores? Isn't just one going to be enough since I will be doing a secondary (agar to agar) transfer anyway? Is there any benefit in creating as many primary agar dishes as possible?
Thanks!
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MonkeyJesusFresco
am i suspended in agar?



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I do it kind'a arbitrarily;
i recommend: making a few (i make 3-4 b/c I can, and i dgaf that's how i roll)
that way, if one contams out or goes fuky on you, there are plenty of back ups, and hell, you may not feel like investing the time and resources in cleaning up your only culture 
and don't let the whole plate colonize, have some class.
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dowodenum
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Registered: 08/05/19
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Sometimes the plate is too contaminated. You might be able to clean it up with enough work, but it'll take a while, and it's not a guarantee.
More plates = better odds = shorter time to fruit. If time is no question and you're trying to conserve the print, just do one and cross your fingers. Of course, opening the print multiple times will decrease its sterility!
I've read that swabbing and burying the swab in the (soft) agar works best, but yet to try that myself.
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fiddle_head
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Most people do 3-4. I do 5-6. I like to swab a few plates the same way you'd isolate an axenic culture away from bacteria, yeast etc. and then chop the tip into two pieces with my scalpel. Then I bury those pieces in the last few plates. If you're just scraping though go for a freshly slame-sterilized inoc loop and cool it in agar before you scrape. In the spirit of increasing our chances for success as much as possible, let's do more than one plate, shall we? Always better to have a few backups if some fail. I just used this tek to germ some RW, but the same principles apply for an inoc loop and a print.
https://www.shroomery.org/forums/showflat.php/Number/28160683
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Home Grower
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Thanks, guys, everything you said makes sense. I will follow your advice and will do more than just one starter dish. I will see how many dishes fit in my pressure cooker and will just go with that number.
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MonkeyJesusFresco
am i suspended in agar?



Registered: 10/09/12
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Quote:
I will see how many dishes fit in my pressure cooker and will just go with that number.
hey... save some for transfers
-------------------- LAGM v 2.024
- endo cabendo
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fiddle_head
fettucine embargo



Registered: 08/05/08
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Quote:
Home Grower said: Thanks, guys, everything you said makes sense. I will follow your advice and will do more than just one starter dish. I will see how many dishes fit in my pressure cooker and will just go with that number.
Well, theres alot of middleground between the two, depending especially on which type of PC you have. I don't use no-pour, but I prep mine 100's at a time and weeks in advance most of the time, so pour is easier. 3-4 germ plates is sufficient to isolate an axenic culture away from any competitor molds or bacteria
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Shrimps
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I always only do two germination plates per culture. Why waste that much? You just need it anyway to get a clean transfer piece. That way you can go with more cultures simultaneously
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Agar - The way to go!
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Recognizing and dealing with contamination
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fiddle_head
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Yeah, it's just preference. In the end it's an odds game. 2 is plenty but for someone just starting out who hasn't gotten their aseptic inoculation technique down pat yet i say what's the harm in 3-4 if it's no extra work. An increased number of germ plates could lend itself to a wider set of scenarios, it's just percentages and likelihoods For example, out of 2 plates 1 never germs. 1 germs but develop trich. 3 Transfers are taken, 1 is weakened by bacteria and displays spiky, erratic growth, 1 is mercurial and doesn't take, 1 shows a distinct sector or 2 but also develops trich. There could have been an additional variable which might have presented as a clean germ plate. When I first started with agar (assuming OP is) I remember doing 10 germ plates. It was mad overkill but I had a ton of interesting genetic material to choose from.
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Shrimps
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Also true ^^ I always had the problem, since I am working with glass petris, I never have enough (around 50-60) so I have to cut it a little down to two dishes or often also just 1 xD
I want to order more dishes, but my Lady isn't fond from that Idea xD
-------------------- I am not crazy, I prefer the term mentally hillarious.
Agar - The way to go!
Clean spawn checklist
Proper Surface Moisture
Recognizing and dealing with contamination
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Home Grower
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Re: How many agar dishes? [Re: Shrimps]
#28188102 - 02/15/23 05:23 AM (1 year, 10 months ago) |
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It turned out that my pressure cooker can accommodate only two Polypropylene cups at a time. Yes, you heard that right, only two. The reason for that is because mine is a small 7-liter pressure cooker and the cups are rather large (110 millimeters in diameter). So I did two sterilization runs, two cups at a time.
Now I have 4 presumably sterile dishes. On two of them I forgot to put tin foil on top of the lids, so water dripped in through the micropore tape vent port. These two are probably going to contaminate. This leaves me with just two truly sterile cups.
Now, the cups are very soft and at the same time they have tight lids that snap tightly. They are a pain in the ass to open without totally deforming the cups. It will be challenge to manipulate them inside of the still air box, but I will try my best to make it work.
If the agar cups do well, I will do a single agar to liquid culture transfer, because this will save me a lot of SAB work (as opposed to agar to agar and then agar to grain).
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KeyMaker
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Registered: 01/03/22
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I only have a 250ml media bottle cause I'm dumb, but I can pour 10-12 plates from a full ass bottle, that will be enough for me to transfer the 4 cultures I have.
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