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Opening Petri dish sleeve inside SAB without stirring up air?
    #26680302 - 05/19/20 04:39 AM (3 years, 8 months ago)

Hello, sorry if this is a stupid question but I'm trying to master my SAB technique after many agar failures.

The point is still air, so when I am opening my Petri dishes inside the SAB, it's creating quite a stir. Should I put scissors in the box and cut them and slide them out as slowly as possible? Open the bottom outside of the SAB while spraying Ozium around? Or open them and leave them out ready then let the air settle? Just worried about contam this way.

I'm writing the process before I do so and this is it so far:

1 drop soap to 500ml hot water
Set towel on lid (my SAB is upside down), spray towel, set cookie rack containing items on the towel, spray the SAB while I'm putting it onto the towel and let the air settle.

Then I have the opening dishes problem. Just want to get this right.


Edited by Hide (05/19/20 05:08 AM)


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Offlinepoisoned
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Re: Opening Petri dishes inside SAB [Re: Hide]
    #26680309 - 05/19/20 04:43 AM (3 years, 8 months ago)

I don't use the lid on the bottom, it just makes the surface more wobbly.

Just make efficient moves, don't move your hands above your work and do it.


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How I do glass dishes


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Re: Opening Petri dishes inside SAB [Re: poisoned]
    #26680312 - 05/19/20 04:45 AM (3 years, 8 months ago)

Thank you very much man, I will work on a towel and rack so the surface is a bit more level. But how about opening the actual plastic sleeve of dishes?


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Offlinepoisoned
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Re: Opening Petri dishes inside SAB [Re: Hide]
    #26680338 - 05/19/20 05:08 AM (3 years, 8 months ago)

Open the sleeve in a SAB. Before you'll open the bottle and start pouring all that air will settle out again.


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How I do glass dishes


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OfflineTheShroomyDude
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Re: Opening Petri dishes inside SAB [Re: Hide]
    #26680350 - 05/19/20 05:16 AM (3 years, 8 months ago)

from what ive read agar is difficult to have success with not using a flow hood. im not as experienced as others here but maybe you could replicate the conditions by opening the plate in a small sterilized room. like a closet or something

could try flipping your sab vertically so it blocks any air from in front of you? just spitballing ideas as im planning to get some more plates soon


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Offlinepoisoned
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Re: Opening Petri dishes inside SAB [Re: TheShroomyDude] * 1
    #26680376 - 05/19/20 05:30 AM (3 years, 8 months ago)

Most people here are doing agar in SABs with great success. There's no way to sterilize a closet tho.

Use SAB as the tek says. Don't try to reinvent a wheel.


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How I do glass dishes


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OfflineTheShroomyDude
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Re: Opening Petri dishes inside SAB [Re: poisoned]
    #26680403 - 05/19/20 05:41 AM (3 years, 8 months ago)

he stated he had failed attempts before, stareatclouds used a vertical sab thought it was pretty neat


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Offlinepoisoned
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Re: Opening Petri dishes inside SAB [Re: TheShroomyDude]
    #26680429 - 05/19/20 06:03 AM (3 years, 8 months ago)

It's obviously used for bags, I fail to see how it would help for petris.

OP, how are you sterilising your agar? Do dishes go bad if they are left uninoculated?


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Re: Opening Petri dishes inside SAB [Re: poisoned]
    #26680473 - 05/19/20 06:46 AM (3 years, 8 months ago)

I think my main problem was that I was using a glovebox before and not being so careful in my technique, as I did actually get a few good plates but mostly contam.

Another issue I believe is that I used beakers without kids covered in foil, now I have media bottles for this run.

I use a PC for 20 mins. On high to get to pressure up, then slightly down and timed from then. Thanks for the answer though!


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Re: Opening Petri dishes inside SAB [Re: TheShroomyDude]
    #26680475 - 05/19/20 06:49 AM (3 years, 8 months ago)

Appreciate the input and ideas man cheers :thumbup:


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OfflineA.k.aM
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Re: Opening Petri dishes inside SAB [Re: Hide] * 2
    #26680490 - 05/19/20 06:56 AM (3 years, 8 months ago)

I’ve always just opened one end out of the box and carefully picked up five at a time to inspect them for breaks and then put them inside.

As long as you don’t crack lids it’s ok.


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Re: Opening Petri dishes inside SAB [Re: A.k.a]
    #26680495 - 05/19/20 07:00 AM (3 years, 8 months ago)

Maybe I'll give this a go as well as inside the SAB, see which works best for me. Thanks mate.


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Offlinepoisoned
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Re: Opening Petri dishes inside SAB [Re: Hide]
    #26680516 - 05/19/20 07:25 AM (3 years, 8 months ago)

You should vent your PC before going ahead. Open the valve on that thing and get a boil going for 10 minutes, then close the valve and start the timer when it starts to whistle.


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InvisibleRoger Clemency
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Re: Opening Petri dishes inside SAB [Re: Hide] * 1
    #26680520 - 05/19/20 07:27 AM (3 years, 8 months ago)

I cut up the side of the Petri sleeve with a scalpel inside the SAB and just peel the plastic sleeve away. And then store them in a gallon ziplock after pouring but before inoculation (in stacks of 10)

What kind of PC do you have? Some smaller ones have a weight for like 12 psi. Also, You have to let the steam vent out for 5-10 minutes before you put the weight on and let it get to pressure. Then you turn the heat down and get it stabilized at the correct pressure. You can start the timer once it’s at pressure but if you’re still adjusting make sure you’re not letting it drop below, go down kinda slowly.


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Re: Opening Petri dishes inside SAB [Re: poisoned]
    #26680621 - 05/19/20 08:36 AM (3 years, 8 months ago)

Oh yeah I do that, sorry. Completely forgot lol. Weight always stays off until there's steam.


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Re: Opening Petri dishes inside SAB [Re: Roger Clemency]
    #26680622 - 05/19/20 08:38 AM (3 years, 8 months ago)

That's a great idea, cheers. I do vent lol just forgot to mention. Thanks mate!


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Re: Opening Petri dishes inside SAB [Re: Roger Clemency]
    #26680627 - 05/19/20 08:43 AM (3 years, 8 months ago)

I used to get plates of Fungi Perfecti because I really liked the Falcon brand they sold (I don't know if they still have them), they never cracked and they came in a sleeve of 20 that you peeled open (like the type of package gauze comes in) and it would have it's own sterile plastic sleeve folded up tight inside. Really convient.

You can cut the bag open outside of your SAB, it's fine- just turn the plates so the lids face down and cut the bottom. Holding them tight move into SAB, flip over, stand up and carefully lift the bag off.


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Edited by mushpunx (05/19/20 08:51 AM)


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