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OfflineMattisfat
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Sterile technique question
    #23868358 - 11/26/16 09:32 AM (7 years, 2 months ago)

I know that you should never put your hands over open plates or jars but is it ok to have them over the lid?

I loosen my lids and keep the plate upside down until i need it so i can just lift the plate off the lid when im working.  I only just noticed that when im working i have been hovering my hand over the underside of the lid - is this a problem? Should i try and move the lid out the way after i pick the plate up?


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Re: Sterile technique question [Re: Mattisfat]
    #23868383 - 11/26/16 09:43 AM (7 years, 2 months ago)

Unless doing bacteriological work the agar should be on the bottom and the lid should be on the top.

It sounds like you have your whole dish upside down so the lid is on the bottom and the bottom filled with agar is upside down on top.


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Offlinekrypto2000
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Re: Sterile technique question [Re: bodhisatta]
    #23868401 - 11/26/16 09:52 AM (7 years, 2 months ago)

I'm confused why the underside of your lid is facing upwards, what's going on here? Are you flipping your lid upside down when you take it off? If so I would not do that, I'd be worried contaminants would land on it and get stuck in any condensation that has formed. If you ever question your sterile procedure though just do a control petri, do whatever you normally do minus actually transferring something to it and that way you can see if anything grows.


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Re: Sterile technique question [Re: Mattisfat]
    #23868402 - 11/26/16 09:52 AM (7 years, 2 months ago)

Quote:

Mattisfat said:
I know that you should never put your hands over open plates or jars but is it ok to have them over the lid?

I loosen my lids and keep the plate upside down until i need it so i can just lift the plate off the lid when im working.  I only just noticed that when im working i have been hovering my hand over the underside of the lid - is this a problem? Should i try and move the lid out the way after i pick the plate up?



Just curious where you came across this technique as it's crazy.  Are you seeing a lot of perimeter contams?  You might invest in some parafilm to keep them sealed.  Not trying to be snarky...


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Edited by Mycolorado (11/26/16 09:53 AM)


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OfflineMattisfat
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Re: Sterile technique question [Re: Mycolorado]
    #23868427 - 11/26/16 10:02 AM (7 years, 2 months ago)

I saw videos of someone who did it - it makes the transfer very quick. I thought it was a common technique.

It was these videos that i took the idea from.
https://www.shroomery.org/forums/showflat.php/Number/21032261/fpart/1/vc/1


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Re: Sterile technique question [Re: Mattisfat]
    #23868436 - 11/26/16 10:06 AM (7 years, 2 months ago)

Well, munch seems to rock it.  That said, it looks like a silly, labor-intensive way to do things...


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Edited by Mycolorado (11/26/16 10:13 AM)


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OfflineMattisfat
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Re: Sterile technique question [Re: Mycolorado]
    #23868465 - 11/26/16 10:15 AM (7 years, 2 months ago)

Is there any better videos to watch for technique?

say i have my plates he normal way up, i take it i need to put my lids down the correct way up but do i need to place them on something that has been wiped with iso or does it not matter?

Im new and im still just cleaning up vendor spawn at the moment so i cant really say how my technique has affected it so far.


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Re: Sterile technique question [Re: Mattisfat]
    #23868840 - 11/26/16 12:18 PM (7 years, 2 months ago)

If using Petri dishes,  crack them open just enough to transfer to (30-45 degrees) without your exacto/scalpel touching anything.  The plate you transfer from isn't as crucial as it has now served its purpose and may be discarded. Work quickly with smooth motions.


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OfflineMattisfat
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Re: Sterile technique question [Re: Mycolorado]
    #23868854 - 11/26/16 12:26 PM (7 years, 2 months ago)

Thanks yeah i get it, what about those wedges that just dont easily come off the scalpel? is there a technique for that?


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Re: Sterile technique question [Re: Mattisfat]
    #23868870 - 11/26/16 12:32 PM (7 years, 2 months ago)

Yeah, that's frustrating. Typically happens after shit carbonizes on the blade so make sure it doesn't have build-up.  You can also angle the blade and just barely slice down into the agar you're transferring to leaving the wedge on top...if that makes sense.  Also, stabbing through the mycelium  can cause it to stick.  I try to slip the blade under the myc into the agar.


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Edited by Mycolorado (11/26/16 12:34 PM)


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Re: Sterile technique question [Re: Mycolorado]
    #23868929 - 11/26/16 12:49 PM (7 years, 2 months ago)

I hold my pasty plates entirely upside down. Crack the lid, set the plate on top of the lid. Grab my myc onto scalpel, add the transfer to the still upside down plate. Then simply snap the plate onto the lid.


I've had fantastic results doing it this way.


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Offlinekrypto2000
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Re: Sterile technique question [Re: Mattisfat]
    #23869007 - 11/26/16 01:13 PM (7 years, 2 months ago)

Quote:

Mattisfat said:
Thanks yeah i get it, what about those wedges that just dont easily come off the scalpel? is there a technique for that?




If going into a jar just flick the shit out of it. If going into agar dig into the receiving plate at an angle and it should wipe off eventually. As the other guy said it's due to the carbonized shit on the blade. An alternative to avoid that would be to get one of those scalpels with replaceable blades and just use a fresh sterile blade every time, no need to flame it and thus no carbonization occurs. That's kind of what I do, but I still reuse my blades and just switch them out when it either gets bad or if it's a really important transfer and I don't want to take any chances.


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OfflineMattisfat
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Re: Sterile technique question [Re: krypto2000]
    #23869187 - 11/26/16 02:25 PM (7 years, 2 months ago)

my scalpel is incredibly coated - think i need a blowtorch instead of my alcohol lamp


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Offlinekrypto2000
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Re: Sterile technique question [Re: Mattisfat]
    #23869346 - 11/26/16 03:18 PM (7 years, 2 months ago)

Coated in what? I think it's just from heat if it's the carbon and rust, it's just from the heat.


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OfflineMattisfat
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Re: Sterile technique question [Re: krypto2000]
    #23869462 - 11/26/16 03:52 PM (7 years, 2 months ago)

yeah i just mean carbon, i didnt realise thats what was making it stick


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Offlinemrmazdarx9
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Re: Sterile technique question [Re: Mattisfat]
    #23869543 - 11/26/16 04:21 PM (7 years, 2 months ago)

Blow torch is always better than alch lamp imo


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Re: Sterile technique question [Re: krypto2000]
    #23869860 - 11/26/16 05:49 PM (7 years, 2 months ago)

Quote:

Mattisfat said:
my scalpel is incredibly coated - think i need a blowtorch instead of my alcohol lamp



Agree with mazda on this...torch is better.  Either way, keep your blade at the top of the blue part of the flame to cut down on soot.  It's also the hottest part.

Quote:

krypto2000 said:
Coated in what? I think it's just from heat if it's the carbon and rust, it's just from the heat.




Agar and mycelium can stick to the blade and can leave carbonized residue on the blade, so it's good to wipe it clean every few plates. 

Quote:

krypto2000 said:

Quote:

Mattisfat said:
Thanks yeah i get it, what about those wedges that just dont easily come off the scalpel? is there a technique for that?




If going into a jar just flick the shit out of it.




This is bad advice IMHO.  I don't think flicking anything anywhere has any place in this art/science.


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Re: Sterile technique question [Re: Mycolorado]
    #23869882 - 11/26/16 05:57 PM (7 years, 2 months ago)

http://images.slideplayer.com/18/5669223/slides/slide_35.jpg

I do something like this, I dip my scalpel in some 91% first to clean it off then flame it red hot so I'm not burning agar residue onto the blade between transfers. I use a baby food small glass jar for that.


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Re: Sterile technique question [Re: bodhisatta]
    #23869896 - 11/26/16 06:01 PM (7 years, 2 months ago)

Beauty!


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