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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: I want to share my Agar Technique. [Re: truskool]
#12991252 - 08/02/10 02:57 PM (14 years, 5 months ago) |
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So back to agar- I feel like the high sides on the jars reduce comtams, anybody else think so? And a plastic lid is nice.
-------------------- You make it all possible. Doesn't it feel good?
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2ndChancesRDivine
I slayed the Jabberwock!



Registered: 04/19/10
Posts: 1,100
Loc: ked up in my mind..
Last seen: 2 years, 1 month
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Re: I want to share my Agar Technique. [Re: Doc_T]
#12991478 - 08/02/10 03:44 PM (14 years, 5 months ago) |
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No idea. I've noticed that mycelium like to grow up the wall though.
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All your RC are belong to me.
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rave420
open minded




Registered: 07/20/10
Posts: 694
Loc: Vancouver Island
Last seen: 14 years, 2 months
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because the walls are always moist. The one thing you can do to prevent this is have good air exchange going to that moisture can evaporate, but then again, you don't want your agar to dry out either.
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we all breathe the same air, drink the same water, and draw our strength from the same giant fireball.
Help me with my collection (trades only)
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cubezoid
Stranger
Registered: 01/25/12
Posts: 63
Last seen: 11 years, 6 months
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Re: I want to share my Agar Technique. [Re: rave420]
#15818700 - 02/16/12 01:16 PM (12 years, 10 months ago) |
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So I stumbled across this post and it looks similar to what my friend was considering doing, yet he thinks it may be possible to take mycelia from fully inoc'd jars and transfer via an RTV port into agar mason jars. Does this seem more sterile? Or would there be too many contams on the top layer of verm in the inoc jars? I think since you wouldn't have to open the jar it could be a good way to clone.
Also, since the needles are hollow, he's considered cloning from a fruitbody by scooping tissue from a cut stem with the needle and injecting it onto the agar through the self sealing port. Ideas on feasibility?
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crimsondrac
Stranger
Registered: 12/09/11
Posts: 493
Last seen: 11 years, 7 months
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Re: I want to share my Agar Technique. [Re: cubezoid]
#15818921 - 02/16/12 01:57 PM (12 years, 10 months ago) |
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Nice necro of a 2 year old thread. Should just start a new one.
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cubezoid
Stranger
Registered: 01/25/12
Posts: 63
Last seen: 11 years, 6 months
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Re: I want to share my Agar Technique. [Re: crimsondrac]
#15819182 - 02/16/12 02:55 PM (12 years, 10 months ago) |
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Yeah you're right, I wanted to try and keep all the info contained in one discussion but I'll start a new topic.
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micki_t


Registered: 07/19/12
Posts: 12
Loc: Northumberland, UK
Last seen: 11 years, 15 days
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Re: I want to share my Agar Technique. [Re: rave420]
#16833573 - 09/12/12 08:24 AM (12 years, 4 months ago) |
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I have been doing very similar to you for my agar work (only cloning so far), and am also having great success.
I mix the agar, put in the little jars, and then pc for 30mins. The lids have a hole in them with tyvek filter, and then oven grade rubber disk with hole to seal lid well and protect tyvek from condensation etc, which are placed on the jars just enough to get a grip, covered with foil, and pc'd. I do use a glove box, and have now made a clean area inside a large cupboard, and this seems to work even better (I'm getting minimal contams:).
On reflection, I will be buying slightly bigger jars next time, as the 1.5oz jars are a little small, and the bigger jars that I have used, are much easier to observe through. I also have some glass petri dishes, which work really well too, although are very expensive.
I don't think that these jars would be practical if I was doing a lot more agar work, but do see them as a perfect and cheap method for the hobbyist to gain good sterile results.
Now a little comment about aseptic technique. When I was at university, studying Biomedical Sciences, we did a lot of aseptic work, especially in microbiology, and this was always performed on open bench, in a far less than sterile lab (equiv to your bathroom), with Bunsen and set up. In this situation it is all about the slight of hand, as you described opening the jar lids to inoculate the agar. Another practice that was invaluable, was working closely to the flame of the Bunsen, as this helped carry upwards and away any contaminants from the petri surface. I have also used a laminar flow hood, in a human tissue culture lab, and cannot deny the advantage that these provide, although they are a professional piece of kit, and certainly not necessary for the hobbyist,
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