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Offline81Renaissance
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Registered: 11/01/06
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Flow Hood Necessary?
    #7960346 - 01/31/08 02:07 PM (16 years, 1 day ago)

Was planning doing some agar based strain isolation, and I've read frequently that a flow hood is necessary, but I was curious if I could successfully do agar culturing in a clean glovebox.
If I don't have a flow hood should I just go straight to a multispore LC and forget the agar?


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Invisiblenovicemycology
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Registered: 07/12/06
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Re: Flow Hood Necessary? [Re: 81Renaissance]
    #7960364 - 01/31/08 02:12 PM (16 years, 1 day ago)

I have about a 65/35 success to failure ratio with agar and a glove box. So yes it can be pulled off quite easily with out a flowhood. Just got to extra measures to ensure a still air atmosphere in and around your work area and just be clean. You should have no problem.


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OfflineRogerRabbitM
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Re: Flow Hood Necessary? [Re: novicemycology]
    #7960560 - 01/31/08 02:57 PM (16 years, 1 day ago)

You can probably improve on those odds substantially by placing the glovebox itself into a small, closed room with no drafts. If carpeted, mist the carpet itself well with water to prevent dust becoming airborne out of it. Spray oust in the air for one minute. In fact, I spray oust in a small bedroom for one minute, with a can in each hand.

Just before working in a glove box, spray the inside with water mist. Use the same mister you use on your mushrooms. Having the water droplets all around the glovebox will attract and hold any contaminants that come in contact with them.

Work very fast. A petri dish should be open no more than a few seconds, five at most. Allow your agar to cool in the filtered bottle you sterilize it in, right up until it's almost too thick to pour. Remember, as the agar cools, it shrinks, and when it shrinks it draws in air to replace the volume, thus it draws whatever else is in the air too, including contaminant spores.

As soon as you pour the dishes, slip them right back into the sleeve or a zip lock bag to cool. Keep them in the sealed bag until you inoculate, at which time you want to immediately wrap with parafilm.

I consider the flowhood mandatory for agar work, but that's mainly because my 40 year old condo building in a wet climate is so mold infested as to be dangerous, and I also consider any less than a 99% success rate to be a failure. I still use my glovebox when isolating wild species, because I don't want the flowhood blowing contaminant spores all over the room.
RR


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OfflineHybridprX
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Registered: 01/29/08 Happy 16th Shroomiversary!
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Re: Flow Hood Necessary? [Re: RogerRabbit]
    #7960574 - 01/31/08 02:59 PM (16 years, 1 day ago)

I do all my agar work in a glove box. Works fine for me.


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Offline81Renaissance
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Re: Flow Hood Necessary? [Re: RogerRabbit]
    #7960629 - 01/31/08 03:10 PM (16 years, 1 day ago)

RR you rock the casbah. Thanks for the advice


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InvisibleDay Tripper
Got a goodreason, for taking the easyway out.


Registered: 01/27/08
Posts: 198
Loc: Rocky Mountains
Re: Flow Hood Necessary? [Re: 81Renaissance]
    #7961008 - 01/31/08 04:45 PM (16 years, 1 day ago)

I agree, RR does in fact rock the casbah.


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