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OfflineEp1429
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Registered: 10/10/18
Posts: 800
Last seen: 28 days, 16 hours
Agar plate transfers
    #25613670 - 11/14/18 06:28 AM (6 years, 2 months ago)

I'm doing my first set of agar plates ATM, and I have two plates inoculated with a B+ MS syringe, two plates inoculated with spores from a print I took off the second flush of a B+ BRF-to-bulk shoebox I have going, and two plates inoculated with rice sized pieces of B+ mycelium from inside the stem of the best performer I had growing in another shoebox that was done with wbs for spawn.

The plates are MEA, and I mixed the agar following the directions on the package for what is supposed to be a 4% agar mix, and after 48 hours, they show no signs of contamination.  They are Saran wrapped like Bod did in his Tek videos, and stored individually in ziploc baggies.

Background out of the way, here are my questions:

1) I should expect to see growth from the cloned plates first, then the spore prints, and finally from the syringe plates last, correct?  I've read about a hundred threads on agar development, and that has been my takeaway thus far.

2) When it comes time to do my transfers, I wipe down the outside of the plates I'm working with using 70% iso, and put them in my SAB.  I leave the lid on the plate that is going to receive the transfer until the last moment, and only crack it open enough to get my razor blade inside, but what about the plate with growing mycelium?  Since I'm cleaning it up via transfer, I can take the lid off the donating plate, correct?  I have really large hands and it is going to be difficult to do the work if I must leave the lid mostly closed and the opening is facing away from me, and towards the back of the SAB.

3) When I make my transfers to the new plates, what should I do with the donation plates?  I was going to do glass plates I could PC, but after study, the ten plates that came in the package didn't seem like they would be enough to do two or three transfers from the original, so I went with a sleeve of plastic plates.  Can these be cleaned and sanitized enough for use again?  For example, can I run them through the dishwasher and wipe them down with 70% iso or peroxide, then use them again?

I'm sure I will have more questions about this if anyone responds, so please expect them.  I know agar is going to be easy when I get everything figured out, I work with bacterial agar plates during my 9-5, but I'm still on the learning curve here for mold growth.

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Invisiblepuff4200
Natural born lever puller

Registered: 09/26/10
Posts: 1,269
Re: Agar plate transfers [Re: Ep1429]
    #25613752 - 11/14/18 07:54 AM (6 years, 2 months ago)

1)yes but the syringe and print will be about the same since you have to wait for the spor s to germinate first to see growth.

2) I never leave any plates open, I open them as I take the price out, close it, open the receiving plate, close it. The longer you leave plates open the more chance for contamination you have.

3) no you cannot resterilize plates. When pouring you open a new sleeve of sterilized plates and pour. If your looking for something more availability and reusable you should look into pasteyplates. Also just so you know alcohol dosent sterilize so that plan in flawed on a couple levels. Plates are cheap.


Also try to shorten the post next time most people won't read a book and they'll skip over your post without answering.

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OfflineEp1429
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Registered: 10/10/18
Posts: 800
Last seen: 28 days, 16 hours
Re: Agar plate transfers [Re: puff4200]
    #25613763 - 11/14/18 08:15 AM (6 years, 2 months ago)

Quote:

puff4200 said:

3) no you cannot resterilize plates. When pouring you open a new sleeve of sterilized plates and pour. If your looking for something more availability and reusable you should look into pasteyplates. Also just so you know alcohol dosent sterilize so that plan in flawed on a couple levels. Plates are cheap.


Also try to shorten the post next time most people won't read a book and they'll skip over your post without answering.




I know that alcohol doesn't sterilize, that's why I wrote sanitize. 

BUT hydrogen peroxide DOES sterilize.

https://www.cdc.gov/infectioncontrol/guidelines/disinfection/disinfection-methods/chemical.html

That's why I asked.  I'd rather order a couple of bottles of 7% hydrogen peroxide than sleeves of Petri dishes.

I will probably end up using the pasty plates later, though.  Not a big fan of weird scientific orders suddenly being shipped to my house.

And finally: If they can read the teks and learn enough about mycology to do this hobby, reading five or six paragraphs (less than a page and hardly a book) shouldn't be too difficult.  Especially if they're looking for answers to their questions.  I've read hundreds of threads about this in the last two weeks, for instance.

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OfflineNosmoKing
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Registered: 01/12/18
Posts: 189
Last seen: 2 years, 10 months
Re: Agar plate transfers [Re: Ep1429]
    #25613915 - 11/14/18 09:52 AM (6 years, 2 months ago)

We wish an H2O2 dunk sterilized.  There are some fancy, schmancy machines that use hydrogen peroxide gas to sterilize, but I sure can't afford one of those.

An at home mycologist cannot sterilize the plastic they use to make those cheap dishes, unfortunately.  Disinfection != sterilization.  Hell, Lysol disinfects.  But you don't have to take our word for it.  Try it yourself.  That's the fun part of having a hobby - you can do things the way you want - not how some corporate bigwig (or anonymous internet user) says.

Edited by NosmoKing (11/14/18 09:56 AM)

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OfflineEp1429
Stranger
Registered: 10/10/18
Posts: 800
Last seen: 28 days, 16 hours
Re: Agar plate transfers [Re: NosmoKing]
    #25614038 - 11/14/18 10:45 AM (6 years, 2 months ago)

I know that you can purchase 35% hydrogen peroxide for use in sterilizing water for human consumption, FDA approved. We occasionally have to do that for the wells people’s cows drink from. The CDC link above says 7% is enough.

Doing this in the small mason jars will end up being the most economical, I think.  I was just curious if there was a system of at-home-sterilization I hadn’t come across. There is a lot of information on this site, it’s entirely possible that a person could read a thousand threads and still miss the needle in the hay stack.

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