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Agar Culture Questions and Pics
    #2106887 -

First of all this is treasure coast culture and it was inoculated with a syringe by AFOAF on 11/3. Its been almost 2 weeks now and theres not much growth. The growth that is there seems to be very cottony and slow and doesent look at all like other pics of cultures on agar hes seen.

First Question - What temp should a petri be stored at for colizanation? Methinks around 70-75 degrees.

Second Question - If trying to isolate a pure strain, what part should be transfered to the new medium? Ive seen people saying that you should transfer the rhizomorphic parts, (which I see none of on this dish) and others saying it doesent matter. AFOAF understands sectoring but when it comes to pointing out the good mycelum to transfer he gets lost, and on this dish it kinda all looks the same.

Third Question - What is the proper way to start work with a flowhood that when not in use just sits in a living room in the corner? Spray work area down with lysol as well as the filter on the flow hood and turn on for a few min before starting work?

Thanks in advance for any help.







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"How doth the little crocodile improve his shining tail, and pour the waters of the Nile on every golden scale! How cheerfully he seems to grin, how neatly spreads his claws, and welcomes little fishes in With gently smiling jaws!"

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Re: Agar Culture Questions and Pics [Re: Deviant]
    #2106933 -

1. i store my dishes between 78-84F and it works for me. i'm not sure what the universal best temp would be.

2. rhizo growth often doesn't show up right away, especially when starting with spores. just pick the best looking growth and transfer it.

3. mist the air with a peroxide solution and spray all your working surfaces down real good. if your flowhood has been gathering dust, clean it and let it run for a while before using it. i don't know if you should spray the filter or not. if you are going to spray the filter, let the fan run for a while to blow off whatever dust and bacteria it can first. then, spray it.


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Anno cock? Is that some kind of Greek liquer? -Geo's all-knowing sex slave
Scat's way better than mud cause it's warm. -Ythan 4/23/07

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Re: Agar Culture Questions and Pics [Re: Deviant]
    #2106940 -

well first off, your not gonna be doing yourself any favors by opening it up like that... =) but I'm presuming you kindof gave up on that. secondly, I've heard a lot about spores really not doing well on agar. I'm not sure if this is on a "scrape it off the dry print" or out of the syringe like you, but from what I have heard it really isn't a great way to go. besides, the whole point of agar work is to CLONE something... either that or to seporate out contaminants... but agar isn't well suited for spores, IMHO. (I could, however, be wrong. =)

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Re: Agar Culture Questions and Pics [Re: schorts]
    #2106958 -

actually, agar works wonderfully for spore germination.

it is just more work starting from spores, instead of tissue.


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Anno cock? Is that some kind of Greek liquer? -Geo's all-knowing sex slave
Scat's way better than mud cause it's warm. -Ythan 4/23/07

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Re: Agar Culture Questions and Pics [Re: eric_the_red]
    #2107368 -

I wouldn't spray the filter. Especially the one you have. There are filters that can take moisture but just not that one. I have my box sitting on my workbench on a piece of plastic. I spray down the plastic with 91% alcohol. 70% may bwe better as it won't evaporate as quickly. And then I let it sit for at least 7 minutes. I then spray the plastic once more and turn on the fan. Anything I bring into the flow I spray with alcohol first. And let the alcohol evaporate in the flow. Everything should be sterile by doing it this way. Also I have found that not having a draft shield you need to stay in the middle of the filter face. Leave about 1" around the edges as some unsterile air from the sides may be pulled in and mixed with your clean air off of the filter. A draft shield would provide 100% coverage to within 1" or so of the farthest edge. Remember also that the most sterile items stay the closest to the filter and the less sterile items away or farthest back from the filter. Your alcohol lamp would be right up front and when opening or just prior to closing something get behind the flame to be the most sterile. That's just how I do things and everyone will have their own way of doing things also.

As far as your culture is concerned it looks like that little patch on the bottom is the most cottony. Cottony is alright if that is all you have. I use differant parts of a print in differant areas of the dish to get the most diverse array of spores. And many dishes too. If you are using plastic dishes then they are not that expensive. And agar really isn't that much either considering the benifits you will reap with isolations.

Hope that helped. Good luck and your FOAF is on the right track for sure.


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"Without deviation from the norm, progress is not possible."
--Frank Zappa--

:laugh:

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Re: Agar Culture Questions and Pics [Re: bonsai]
    #2107399 -

Thanks for the help everyone


--------------------
"How doth the little crocodile improve his shining tail, and pour the waters of the Nile on every golden scale! How cheerfully he seems to grin, how neatly spreads his claws, and welcomes little fishes in With gently smiling jaws!"

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Re: Agar Culture Questions and Pics [Re: Deviant]
    #2112973 -

Take it from these areas. There appears to be some sectoring. You should start to really see rhizo after this transfer. Good Luck.



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"Ekam Sat Vipras Bahudda Vidante"
The One Truth is named by the wise in many ways.

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Mushrooms, Mycology and Psychedelics > Advanced Mycology
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