Couple of agar transfer questions
Posted 23 July 2020 - 06:33 AM
1. Why do you transfer tiny bits, instead the whole big chunk and wouldn't it colonize the plate faster? - I answered "because contams", figuring that a small wedge is more likely to be clean and less likely to pick up something along the way. Also, I know that we're supposed to transfer the fastest growing sector, but in this case the growth was uniform. Also, I don't care about isolating a monoculture.
2. Why are you doing only two transfers instead of transfering to all the plates you have? - again, I said "cuz contams, dude", thinking that the longer I have the original plate open, the more likely it is to pick up contams and that the more transfers I do, the less likely they are to succeed. He seemed to think that I shouldn't worry about it and that the more transfers I do, I'll get a clean plate faster.
So, what do you think? How many transfers from one contaminared plate should I aim for and how big of a wedge should I go for?
- Arathu likes this
Posted 23 July 2020 - 08:49 AM
Then I have a question. Why are you doing transfers? Because of contams?
Using a small piece gives you better chances of isolation a single sector or substrain. Which besides trying to clean up a contaminated culture would be the most popular reason for doing transfers at all. Still if chasing a clean culture, a small sample is less likely to have as much contamination as a large sample. Like if you took a whole dish and transferred to it to another dish you are literally transferring all the contamination and substrains to the next plate. A small speck of mycelium is all that is needed to start another dish. Since you are not interested in an isolated strain then maybe you could have answered your friend "Because its cool and makes me feel like a mad scientist".
For question number 2, I don't know why you are only doing two transfers lol. I will say that if you have a 50% success rate and you do two transfers, then you would end up with one good plate. if you did 10 transfers then you would have 5 successful plates and so on. Your success rate is gonna be based on how clean your method is, your understanding of how contams happen and even down to every movement you make while doing transfers. Well planned, smooth and methodical movements in an open air situation can yield a better success rate that haphazardly clumsy shaky movements in a SAB. Every time you get set up and do agar transfers you are essentially practicing and building muscle memory. Which is important for a better future success rate. Then if you only need to or two plates then maybe I would have answered" Dude, I don't need that many plates". But he does seem to grasp the idea well, perhaps invite the buddy to participate in the next transfer session?
Edited by coorsmikey, 23 July 2020 - 08:51 AM.
- mushit, Arathu, PJammer24 and 2 others like this
Posted 29 July 2020 - 12:09 PM
No offense is intended in what Im about to say but it sounds like you do not understand much about agar and what it is useful for and why, Mr Magic.
It's alright, we all must start and learn and experience, no matter no worries.
1. Mikey done said it. He say thing. It true. Less genetics for isolation and also less material for contamination to be passed.
2. You should consider a plate that has been transferred from TRASH. Transfer to as many plates as you want, but 2 is too few. At least 5 transfers would be a useful number to hedge and pick good growth from for the next steps.
But then it comes to ... why are YOU using agar Mr MAgic?
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Posted 02 August 2020 - 08:56 AM
I seriously recommend getting a copy of "Growing Gourmet and Medicinal Mushrooms" by Paul Stamets.......
Agar work is a collection of items, media, and techniques that allow us the possibility to achieve cultivation goals such as:
- Spore germination
- Clone propagation
- Isolation of species (from "contaminants")
- Isolation of strains
- Propagation of particular phenotype's
- Evaluation of a species and strain(s) for growth/fruiting characteristics
- Training of species and strain(s) to consume particular substrates and substances
- Long term storage of particular cultures via slants thus enabling the building of culture libraries
I'm sure there are more reasons to use these tools, YOU as the cultivator, decide the why(s) and the how(s) you'll use them.....
This is a great book, IMHO, to have in your library....
Note: I burn through sleeves of plastic pitri plates to the point that I'm moving towards all glass (of course as I can afford to do that)
There's enough plastic pollution that it's a permanent component of the biosphere now.....IMHO
Good vibes coming atcha friend.....
Edited by Arathu, 02 August 2020 - 09:16 AM.
- ElPirana likes this
Posted 02 August 2020 - 10:11 AM
The smallest piece, tissue and medium, transferred from one plate to another the better the odds of beating contaminants/accomplishing some other of the above goals. For example shooting for strain isolation..especially with good aseptic technique..If that's what you're doing/trying to accomplish, however, you can also transfer larger slices of colonized agar plates to rapidly expand a particular culture once isolated and demonstrating desirable characteristics (we have to figure out what desirable characteristics means as related to the organism), especially in preparation for a scale grain spawn run of it. That's the next step in fact, expansion....I cut oyster pitri plates like a pie when transferring to grain. I'm trying to do the same with Maitake but just haven't found vigor yet, I want to fruit them in half barrels full of dirt and sawdust......I'm missing something.
How many strains are here? Is there bacterial infection on this plate? See anything of interest or worth further isolation and growing? How would we know? What transfers should be made?
Isn't learning fun?!